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NB100-304

53BP1 Antibody - BSA Free

(14 Reviews)

(712 Publications)

0.1 mg

NB100-304

Irradiated cochlear spiral ganglion cells, mouse, frozen section of fixed material. Image from verified customer review.

<b>Genetic Strategies Validation. </b>53BP1 was detected in immersion fixed HeLa cells (left) but was not detected in 53BP1 knockout Hela cells (right) using Rabbit Anti-human 53BP1 polyclonal antibody (Catalog #NB100-304) at 0.3 ug/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to nuclei.

Total protein from HeLa, A431, Neuro2A, and PC12 was separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 1.0 ug/mL in 5% block buffer and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. The observed molecular weight for these samples are ~250 kDa and the theoretical molecular weight is 214 kDa.

Human breast tumors stained with 53BP1 antibody. Image from verified customer review.

Detection of Human and Mouse 53BP1 by IHC. Sample: FFPE sections of human ovarian carcinoma (left) and mouse teratoma (right). Antibody: 53BP1 Antibody (Catalog #NB100-304) used at a dilution of 1:1000 (1ug/mL). Detection: DAB.

Human medulloblastoma (DAOY) and mouse astrocyte (C8D1A) cell lines were exposed for 48 hours to DMSO or 1ug/mL of the DNA damaging agent EPE. Cells were immunostained for 53BP1 (green). The nuclei were counterstained with DAPI (blue). Image from verified customer review.

An intracellular stain was performed on RH-30 cells with 53BP1 Antibody (Catalog #NB100-304AF647) (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Immunoprecipitation analysis lysates from HCC44 cells in 1% NP40. Image from verified customer review.

<b>Biological Strategies Validation. </b>Effect of RK-33 on radiation-induced DNA damage. A Immunofluorescence images showing 53BP1 and gamma-H2AX foci in A549 cells after 2-Gy radiation and A549 cells pre-treated with 6 uM RK-33, 12 h before radiation. Overlap of 53BP1 and gamma-H2AX is seen in the merged picture of the co-immunofluorescence staining. Scale bar is 2 um. Image collected and cropped by CiteAb from the following publication (//embomolmed.embopress.org/cgi/doi/10.15252/emmm.201404368) licensed under a CC-BY license.

Lamin A/C-53BP1 interaction is regulated in a DNA damage-dependent manner. U2OS/GFP-lamin A cells were pretreated with 10 um ATMi f for 1 h before exposure to IR (10 Gy, 1 h recovery). Cell extracts were then subjected to immunoprecipitation using GFP-Trap beads, and bound complexes were then analyzed by immunoblotting using 53BP1 and GFP antibodies. WCE represents 1% input. Image collected and cropped by CiteAb from the following publication (//doi.wiley.com/10.1111/acel.12258), licensed under a CC-BY license.

HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with 53BP1 Antibody conjugated to DyLight 550 (NB100-304R) at 5 ug/ml for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

An intracellular stain was performed on Ntera2 cells with 53BP1 Antibody NB100-304 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Whole cell lysate from U2OS or 293T cells. Bands indicate an observed molecular weight of ~220 kDa and the theoretical molecular weight is 214 kDa.

<b>Biological Strategies Validation. </b>Upper Panel: 53BP1 foci in proliferating MEFs. Lower Panel: 53BP1 foci in proliferating MEFs exposed to 10 Gy of IR.

Embryonic Fibroblast cells pre-extraction for 5 mins with CSK buffer. Fixed with 4% PFA and 75% Ethanol. Primary Antibody at 1:1000. Secondary Antibody at 1:1000. Image from verified customer review.

<b>Biological Strategies Validation. </b>Upper Panel: Control untreated cells. Lower Panel: Cells exposed to irradiation (10 Gy) and probed for 53BP1 foci. Cells were grown on coverslips, fixed with 4% paraformaldehyde, methanol permeabilized, blocked for 1 h, RT. Incubated with primary antibody (1:200) overnight, washed 3x with PBS, probed with tubulin (Alexa Fluor 594) antibody for 2 h, RT. Washed 3x with PBS, mounted on slides using prolong gold, imaged using Nikon confocal microscope (100x oil). Image from verified customer review.

<b>Genetic Strategies Validation. </b>MUS81 inhibition in BRCA2-deficient cells causes accumulation of 53BP1 nuclear bodies and G1 arrest. H1299 cells carrying a DOX-inducible BRCA2 shRNA were transfected with control or MUS81 siRNAs. Representative images of cells processed 72 h later for immunostaining with 53BP1 Antibody (green) and anti-cyclin A (red) antibodies. DNA was counterstained with DAPI. Scale bar, 10 um. Image collected and cropped by CiteAb from the following publication (//www.nature.com/doifinder/10.1038/ncomms15983), licensed under a CC-BY license.

<b>Biological Strategies Validation. </b>Pot1a prevents DDR in HSCs. Telomeric DDR in 8 week-old LSK cells upon Pot1a knockdown. Immunocytochemical staining of TRF1 (green). Foci co-stained with TRF1 and 53BP1 were identified as TIFs. Nuclei were stained with TOTO3 (blue). Scale bar, 2 um. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41467-017-00935-4), licensed under a CC-BY license.

Lamin A/C-53BP1 interaction is regulated in a DNA damage-dependent manner. U2OS cells were transfected with siCTRL or siLMNA and subjected to laser micro-irradiation, fixed 15 min later and then processed for immunofluorescence with gamma-H2AX and 53BP1 antibodies. Scale bar, 10 um. Image collected and cropped by CiteAb from the following publication (//doi.wiley.com/10.1111/acel.12258), licensed under a CC-BY license.

Ntera2 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-53BP1 Antibody NB100-304 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:1000 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

NIH3T3 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-53BP1 Antibody NB100-304 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:1000 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Staining of 53BP1 in human colon cancer using DAB with hematoxylin counterstain.

An intracellular stain was performed on A431 cells with 53BP1 Antibody (Catalog #NB100-304AF647) (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

For 53BP1 staining, samples were incubated for 1 hour with 1:200 53BP1 primary antibody in PBS/BSA 1%, at room temperature. Green: 53BP Blue: DAPI. Image from verified customer review.

DYNLL1 is required for 53BP1-dependent p53 responses to Nutlin-3. a Immunoblot analysis of the MCF-7 cell lines used in (a) with anti-53BP1 or anti-RPA32 antibodies prior to N3 treatment. b Quantification of n = 3 independent experiments represented in (a), each performed in triplicate. Mean ± SD. c Indicated parental or stably complemented 53BP1-/- MCF-7 cell lines were incubated in the presence (11 days) or absence (7 days) of Nutlin-3 (4 µM)

Depletion of PNUTS and NPAT induces a global DDR.(A) Induction of telomere dysfunction–induced foci (TIF) upon candidate knockdown in HeLa cells from Fig 5. Depletion of TRF2 was used as a positive control. 53BP1 was detected by indirect IF and telomeres by FISH with a Cy3-[CCCTAA]3 probe. (B) Frequency distribution of the number of TIFs per cell from (A). n indicates the number of cells analyzed for each condition and red lines mark the mean.

HPV+ HNSCCs have decreased expression of NHEJ and HR proteins including DNA-Pk and BRCA2Cells were treated with mock or 4 Gy radiation, harvested and lysed at 15 minutes post-treatment, and analyzed by western blot for relative expression of indicated proteins. beta -actin was used as a loading control. Shown is a representative blot from 2 independent experiments.

LNCaP cells in culture were treated for 24 hrs with DMSO or ENZ, then irradiated with 0 or 1 Gy.Cell were fixed and immunostained for 53BP1 0.5, 1.5 and 24 hrs after irradiation. Immunofluorescent images were obtained by confocal microscopy. White bar = 20 microns. A) DMSO alone, B) ENZ alone, C) 1 Gy, D) ENZ + 1 Gy. White bar = 20 microns.

Representative immunohistochemistry staining for 53BP1 expression in pancreatic adenocarcinoma. (A) is high intensity of 53BP1 expression and (B) is low intensity of 53BP1 expression.

Absence of Roma Leads to Activation of the DNA Damage Response and Genomic Instability(A) Representative immunofluorescence analysis of RPA2-pT21, gammaH2AX, and 53BP1 foci in tissue sections from WT and Roma-/- glands at 10dL. The white arrow indicates nucleolar gammaH2AX. Scale bars, 10 um. The higher magnification panels on the right have been chosen to highlight the foci and are not zoomed images of the left panels. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27149840), licensed under a CC-BY licence.

Persistent DNA damage exists in ALT cells. (A) IF-FISH showing DNA damage response (53BP1) in ALT (VA13, U2OS, SAOS2, SKLU-1), telomerase positive (MCF7, A549), and human normal fibroblast cells (MRC5, BJ). Scale bar: 10 um. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31115790), licensed under a CC-BY licence.

Persistence of repair foci at suboptimal pHe. (A) Left panel shows immunofluorescence detection of gammaH2AFX and 53BP1 following 24 h recovery from bleomycin treatment. Control, sample before damage induction. Colocalization is shown in white; PCC, Pearson correlation coefficient score. Right panel corresponding to gammaH2AFX foci/cell at 24 h and 48 h following recovery. Only foci larger than 10 pixels were counted. Experiments were performed in triplicate. Values are mean +/- SEM. *P 3 (c) gamma-H2AX or d 53BP1 foci per nucleus. n = 90 cells from three independent experiments. Mean +/- SEM; two-way ANOVA (gamma-H2AX: F = 35.77, DFn = 4, DFd = 20, p 10 foci was called positive and counted). DAPI was used for nuclear staining. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28489060), licensed under a CC-BY licence.

USP15 regulates homologous recombination (HR) and breast cancer response to poly-(ADP-ribose) polymerase (PARP) inhibitor. c) Non-homologous end-joining (NHEJ) efficiency in USP15-depleted cells were determined using the EJ5-GFP reporter assay, as described in the Methods section. CtIP or 53BP1 small interfering RNAs (siRNAs) were used as a positive control, respectively, in b, c. Data are shown as average of three independent experiments, presented as mean +/- SD, two-tailed Student's t test, *P < 0.01. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30874560), licensed under a CC-BY licence.

MRE11 poly(T)11/12 sequence variant has no apparent functional consequence on DNA damage detection and repair. (D) DNA repair foci was observed by immunofluorescence after 1.5 Gy IR followed by 1 h and 4 h recovery. Irradiation resulted in the formation of both MRE11 and 53BP1 foci in Daudi and Raji cells, whereas Jurkat failed to form MRE11 foci. Nuclei were counterstained with DAPI. Merged figure presents co-localization of MRE11 and 53BP1 foci visualized in yellow. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/21637783), licensed under a CC-BY licence.

Phenotypic consequences of R325Q mutation. (D) DNA damage is increased in patients' B-LCLs as compared to control, with an accumulation of 53BP1 foci. Left: immunostainings of 53BP1 imaged by confocal microscopy, Right: quantification of 53BP1 foci number. Graph represents the average count obtained from two independent experiments (Mean +/- SEM). Total number of cells in control = 508; P2_E71K n = 315; P_R325Q n = 452; Sidak's multiple comparisons test, ****p < 0.0001. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31787977), licensed under a CC-BY licence.

Bystander signaling evokes DNA damage foci formation in urothelial cancer and normal urothelial cells. (A) Schematic representation of experimental conditions including non-irradiated, uniformly-irradiated and shielding protocols where 50% of the slide was exposed and 50% shielded with MCP alloy. Cells were fixed and processed for 53BP1-immunofluorescence, one hour following irradiation. Micrographs of urothelial cancer T24 cells labelled with anti-53BP1 (green) to label nuclear DNA double stand break foci and DAPI to counterstain nuclei are shown for each experimental condition. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29228614), licensed under a CC-BY licence.

Cry2-/- cells accumulate damaged DNA.(A) Representative early passage (P3-4) primary wildtype (WT), Cry1-/-, Cry2-/-, and Cry1-/-;Cry2-/- adult ear fibroblasts stained with anti-53BP1 antibody (green) and DAPI (blue). Insets show enlarged view of indicated cells. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25756610), licensed under a CC-BY licence.

Chromatibody-driven DDR alteration. (B) RNF8-dependent H2A di-ubiquitylation. HT1080 cells were transfected with the indicated constructs for 24 h. Whole-cell extracts were analyzed with an anti-ubiquitinylated H2A antibody (UbH2A). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/27206857), licensed under a CC-BY licence.

The susceptibility of human LECs to low-dose ionizing radiation. The human lens cell line FHL124 was exposed to low-dose IR up to 2.28 Gy. Exposed cells were then processed for both immunofluoresence microscopy (a) and immunoblotting (b) 1 h later. The signals obtained by immunoblotting were quantified and the mean from three independent experiments calculated and plotted (c) against IR dose. GAPDH was used as a loading control. Both gammaH2AX and RAD51 increased linearly with IR dose. Signals for other markers of DNA repair, 53BP1, MRE11 and TP53, remained unchanged as assessed by immunoblotting (b). By immunofluoresence microscopy (a), MRE11 and 53BP1 redistributed into nuclear foci, particularly at the 2.28 Gy level. TP53 remained uniformly distributed throughout the nuclear compartment, but excluded from nucleoli (a). As the levels of gammaH2AX and RAD51 increased after exposure to IR (c), so the number of nuclear foci also increased (a). Scale bar, 10 um. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25924630), licensed under a CC-BY licence.

General chromatin effects of Adeltarod and Adeltaroddeltahbs. (c) No co-localization between Adeltarod structures and 53BP1 in HeLa cells indicates that chromatin is not damaged by the structures. Image collected and cropped by CiteAb from the following publication (//www.mdpi.com/2073-4409/6/2/9), licensed under a CC-BY licence.

Specificity of antibodies for tryptophan 2,3-dioxygenase (TDO) by Western blot (A) and immunofluorescent staining (B). A: The total protein (20 μg) of the dentate gyrus, extracted from the wild-type (lanes 1 and 3) and Tdo-/- mice (lane 2), was subjected to 4% to 12% gradient sodium dodecyl sulfate polyacrylamide gel electrophoresis, and transferred to polyvinylidene difluoride membranes. TDO was detected as described in the Methods section. Note that a 45-kDa band was detected with the anti-TDO antibody. The positions of molecular weight markers are shown on the left. Lane 3: negative control without the primary antibody. beta -Tubulin is a positive control for the Western blot. B: The hippocampal sections of wild-type and Tdo-/- mice were stained with anti-TDO antibody. In sections of Tdo-/- mice, immunofluorescence signals were not detected in granule cells, interneurons (arrowhead), CA1, or CA3 cells, suggesting that the secondary antibody used in this study bound specifically to the primary antibody (mouse IgG). g, granule cell layer; h, hilus. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/20815922), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$CO attenuates mitochondrial dysfunction-derived sterile inflammation through TFEB-mediated mitophagy and mitochondrial biogenesis.Mice were exposed to LPS/D-galactosamine (D-GalN) with or without CO (250 ppm, 2 h/day) pretreatment, the blood and liver samples were collected 2 h and 6 h (b–i) after LPS/D-GalN administration. a The levels of ALT and AST were determined. Data are shown as mean ± SEM (n = 5). ***P < 0.001. b Liver sections were stained with H&E for morphological evaluation. c The levels of serum IL-1 beta were measured by ELISA. Data are shown as mean ± SEM (n = 4). **P < 0.01. ***P < 0.001. d Cytosolic mtDNA amounts in liver tissues were measured by qRT-PCR. Data are shown as mean ± SEM (n = 4). *P < 0.05. e Cleaved caspase-1 in cell lysates isolated from liver tissues was analyzed by immunoblotting. f The level of nuclear TFEB in liver tissues was determined by immunoblotting. Lamin B served as the nuclear standard. g Mitochondrial fractionation in liver tissues was analyzed by immunoblotting using antibodies against PINK1, Parkin, and LC3B-II. COX IV was used as marker for mitochondrial fractions. h, i The levels of autophagy-related genes and lysosomal genes (h) or mitochondrial biogenesis-associated genes (i) expressed in liver tissues were analyzed by qRT-PCR. Data are shown as mean ± SEM (n = 4). *P < 0.05, **P < 0.01. ***P < 0.001. j A scheme for the mechanisms of attenuating LSP/D-GalN-induced liver failure by CO through maintaining the balance between mitophagy and mitochondrial biogenesis. CO/CORM inhibits complexes of the mitochondrial respiratory chain (1) and then induces mitochondrial ROS (mtROS) production (2). Increased mtROS leads to PERK activation in the mitochondria-associated membrane (MAM) (3). The phosphorylation of PERK by mtROS increases intracellular calcium concentration. The activation of calcineurin by increased [Ca2 + ]i causes dephosphorylation of TREB (4), followed by its translocation into the nucleus. The nuclear translocation of TFEB increases the expression of genes involved in mitophagy (5) and mitochondrial biogenesis (6) Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30333475), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Precision quantitative image analysis workflow for the characterization of functional ITH.(a) Representative areas with positive immunohistochemical (IHC) staining for HIF-1 alpha , HIF-2 alpha , phospho-mTOR S2448, phospho-S6RP S235/236 and Ki-67. Only nuclear HIF-1 alpha and HIF-2 alpha expression was included in the analysis. Scale bar, 100 μm. (b) Overview of the quantitative IHC analysis process using a representative image of a whole-slide scanned pT1M1 tumour after phospho-mTOR S2448 staining. The image was overlaid with a virtual grid consisting of 1 mm2 squares that defines the tumour regions to be analysed. Non-cancerous tissue was subtracted manually (dark grey) and a positive pixel count was performed on the corrected area. Positivity per square was expressed as percentage of positive pixels. If a square contained non-tumorous tissue, the positivity was normalized to the corrected square area. Computed values were depicted as a heat map. Coloured squares represent tumour, whereas open squares indicate completely non-tumorous areas. Squares defined as peripheral tumour zone are marked by a black line. Scale bar, 1 cm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/27291893), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reduction of HIF-1 alpha and HIF-2 alpha levels leads to differential effects on apoptosis in RCC10 cells. (A) RCC10 cells stably expressing wild-type VHLp30 or expressing a control shRNA targeting luciferase (Luc shRNA) or a pool of two shRNAs targeting either HIF-1 alpha or HIF-2 alpha were created. These cells were grown to confluence and lysed. Cell lysates were equally loaded and separated by SDS-PAGE. Western blots were performed for VHL, HIF-1 alpha , HIF-2 alpha , GLUT-1, and alpha -tubulin. (B) The RCC10 cell lines described in (A) were grown to confluence and treated with ultraviolet (UV) light and lysed one day later. Cell lysates were equally loaded and separated by SDS-PAGE. Western blots were performed for PARP, cleaved caspase-3, and alpha -tubulin. (C) The RCC10 cell lines described in (A) were glucose starved for 24 hours as described in Methods. Cell lysates were equally loaded and separated by SDS-PAGE. Western blots were performed for PARP, cleaved caspase-3, and alpha -tubulin. (D) The RCC10 cell lines described in (A) were serum starved for 2 days as described in Methods. Cell lysates were equally loaded and separated by SDS-PAGE. Western blots were performed for PARP, cleaved caspase-3, and alpha -tubulin. Similar results were seen with serum starvation for 4 and 6 days (data not shown). Image collected and cropped by CiteAb from the following open publication (//cancerci.biomedcentral.com/articles/10.1186/s12935-015-0175-3), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Expression of hypoxia-inducible factor (HIF) proteins after bilateral renal ischemia or sham ischemia. Immunoblots for HIF-1 alpha (A–C) and HIF-2 alpha (D–F) of tissue extracts from the cortex and outer and inner medulla of the left kidneys of rats 24 h and 5 days following recovery from either sham ischemia (○) or bilateral renal ischemia (●); n = 6 per group. G: typical image of the gel following electrophoresis. H: typical image of the nitrocellulose membrane following transfer. Values are expressed as medians (25th percentile, 75th percentile). Paired comparisons were performed using the Mann-Whitney U-test. Because paired comparisons were made at two time points, P values were conservatively adjusted using the Dunn-Sidak method with k = 2. PTr, PT, and PTr*T are the outcomes of two-way analysis of variance on ranking with the factors treatment (Tr) and time (T). AU, arbitrary unit; I1, 24 h after ischemia; I5, 5 days after ischemia; S1, 24 h after sham ischemia; S5, 5 days after sham ischemia. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30110566), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Active DNA end processing in late micronuclei compartments. a: Detection and quantification of RPA32 phosphorylation (pSerine 33) and ssDNA by specific antibodies in MN compartments of OVCAR-8 cells 36 h after 2-Gy irradiation. Note the nuclear signals have reduced while those in MN were intense. n = 3 biological repeats. Error bars = s.d. **P < 0.01; ***P < 0.005 (t-test). b: Immunostaining and enumeration of phosphorylated NBS1-pSerine 343, BRCA1-pSerine 1524 and TP53-pSerine 15 in MN compartments. **P < 0.01; ***P < 0.005 (t-test). c: Representative images (Upper panel) and quantification of RPA signal in MN compartments (Lower left) or RPA foci in nuclei (lower right) of post-IR OVCAR-8 cells after indicated treatments. Pro-resection genes (MRE11, BRCA1 and WDR70) were knockdown by specific siRNA. Percentage of positive RPA32-pS33 MN population was calculated after 36 h of IR treatment Image collected and cropped by CiteAb from the following open publication (//bmccancer.biomedcentral.com/articles/10.1186/s12885-018-4347-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Curcumin Analogs Induce Apoptosis in Cancerous Cells by Several Pathways. (a) E6-1 (Jurkat), dominant negative FADD (dnFADD) Jurkat, and overexpressing BCL-2 Jurkat were treated for 48 hours then stained for Annexin V and PI (b) E6-1 cells were plated and treated with or without the broad spectrum caspase inhibitor ZVAD(oMe)-FMK for 48 hours. Cells were stained for Annexin V and PI. Results were obtained using image-based cytometry with the Y-axis representative of percent of cells positive for Annexin V (green), PI (red), Annexin V and PI (yellow), or negative for both Annexin V and PI (blue). Values are expressed as a mean ± SD from three independent experiments. (c) E6-1 cells were treated for 24 hours with or without the broad spectrum caspase inhibitor ZVAD(oMe)-FMK and the studied compounds, lysed and subjected to Western blot analysis. (d) NHF and NCM460 cells were treated for 48 hours and 72 hours respectively, lysed and subjected to Western blot analysis. Bands were visualized with a chemiluminescence reagent. Images are representative of three independent experiments. Statistical calculations were performed using Two-Way ANOVA multiple comparison. *p < 0.05 vs % viable of Control (DMSO); #p < 0.05 vs % viable cells for groups without Z-VAD(oMe)FMK. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28439094), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

PEM induces apoptosis of NSCLC cells via YIPF2-TNFRSF10B axis.a, b A549 (a) and H1792 (b) cells were treated with PEM at 5.0 μM for the indicated times (0, 6, 12, 24, 36 and 48 h). Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B and ACTB. c A549 cells were treated with doxorubicin (DOX) at various concentrations (0–2.0 μM) for 18 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B and ACTB. d Overexpression of YIPF2 in H1792 and H1299 cells in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B and ACTB. e Knockdown of YIPF2 expression by YIPF2–1 siRNA in A549 and H1792 cells in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B and ACTB. f Overexpression of YIPF2 in H1299 cells (left) or Knockdown of YIPF2 expression by YIPF2–1 siRNA in H1792 cells (right) in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10A and ACTB. g Three A549 cell lines (Ctrl, YIPF2, YIPF2 + siTNFRSF10B) in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B, CASP8, CASP3 and ACTB. h Three H1299 cell lines (Ctrl, siYIPF2–1, siYIPF2–1 + TNFRSF10B (short isoform)) in the presence or absence of PEM at 5.0 μM for 36 h. Cell lysates were analyzed by Western blotting with antibodies against YIPF2, TNFRSF10B, CASP8, CASP3 and ACTB. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32303681), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Inhibition of CIP2A by siRNA impeded cell viability and DNA synthesis in HPV‐16E6–expressing cells. A, Elevated expression of CIP2A protein in 16E6‐expressing RPE1 cells. B, Western blot analysis of CIP2A and p53 proteins after transfection with scrambled siRNA (siCon) or CIP2A siRNA (siCIP2A) for 48 h. A representative of 3 independent experiments is shown. C, Cell viability assay of RPE1‐16E6 cells with CIP2A knockdown. D, Representative flow cytometry of BrdU staining profiles is shown. E, The mean and SD of BrdU‐positive cells from 3 experiments are summarized. **, P < .01. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29893470), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Characterization of CIP2A expression in a panel of normal human melanocytes (NHM) and melanoma cell lines. (A) Immunoblot analysis and (B) quantification of protein expression levels of CIP2A (C) CIP2A mRNA expression measured by qRT-PCR. NHM1 line was used as calibrator. (D) Cytoplasmic and nuclear expression of CIP2A in normal melanocytes, primary and metastatic melanoma cell lines assessed by immunoblot analysis and (E) by immunohistochemical staining. CIP2A, cancerous inhibitor of protein phosphatase 2A. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/25663244), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$TAZ induces Tfam protein expression at the translational level.a The expression of mitochondrial biogenic factors, including Tfam, Ppargc1a (Pgc1 alpha ), Nrf1, and Nrf2, was analysed using quantitative reverse transcription (qRT)-PCR in the gastrocnemius muscle of wild-type (WT) and muscle-specific TAZ knockout (mKO) mice; n = 6 for each condition (Tfam; *p = 0.0498, Nrf1; *p = 0.0467). b Protein was analysed via immunoblotting to detect TAZ and the indicated mitochondrial biogenic factors in (a). Alpha-tubulin was used as the loading control. c RNA was isolated from the control (Con) and TAZ knockdown (KD) C2C12 myotubes, and the expression of the indicated mitochondrial biogenic factors was analysed using qRT-PCR. The experiments were performed in triplicate (Tfam; *p = 0.04). d Protein from the Con and TAZ KD C2C12 myotubes was assessed via immunoblotting to detect TAZ and mitochondrial biogenic factors. Beta-actin was used as the loading control. e RNA was isolated from WT, TAZ-knockout (KO), and TAZ-rescued (T) mouse embryonic fibroblasts (MEF), and the indicated mitochondrial biogenic factors were analysed via qRT-PCR. f Protein from WT, KO, and T MEF was assessed via immunoblotting to detect TAZ and the indicated mitochondrial biogenic factors. Beta-actin was used as the loading control. g Protein was isolated from the gastrocnemius muscle of WT and mKO mice and analysed via immunoblotting to detect TAZ, YAP, and proteins involved in the translation of both phosphorylated and total forms. Alpha-tubulin was used as the loading control. h Protein obtained from the Con and TAZ KD C2C12 myotubes was assessed via immunoblotting to observe TAZ and proteins involved in translation. Beta-actin was used as the loading control. i Protein was isolated from WT, KO, and T MEF and analysed via immunoblotting to detect TAZ and proteins involved in translation. Vinculin was used as the loading control. j The polysome profile of cells described in panel i was evaluated continuously by measuring absorbance at 254 nm. The 40S, 60S, 80S, and polysome fractions are denoted by the corresponding peaks. k The distribution of Tfam, Atp5d, and Gapdh mRNA from cells described in (j) across a density gradient was analysed using reverse transcription (RT)-PCR. bp = base pairs. For a, b, g 8 to 10-week-old mice were used. Data are presented as mean ± SEM for (a), mean ± SD for (c, e). Statistical significance was analysed via two-sided t-test for (a) and one-sided t-test for (c). One-way ANOVA with Tukey’s multiple comparison test was used for (e). Representative data was shown and experiments were performed at least twice with similar results for (b, d, f–i, k). Source data are provided as a Source data file. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35115527), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Dynamics of the hypoxia-triggered decrease in MET phosphorylation and its reversal upon reoxygenation(A) MCF10A cells were incubated under normoxia or hypoxia for 1 h. They were then treated, under the same oxygen pressure, with 10 ng/mL HGF/SF for 5, 10, 20, 30, 60, 90 and 120 minutes. A control (Ctrl) without any HGF/SF stimulation was also performed. The same amount of protein was analyzed by western blotting with antibodies directed against: phosphorylated residues in the MET kinase domain, the MET kinase domain, the hypoxia marker HIF1a, phosphorylated Akt, Akt, Erk2, phosphorylated Erk, and actine. The positions of prestained molecular weight markers are indicated. Arrows indicate the positions of precursor and mature full-length MET. (B) MCF10A cells were incubated under hypoxia for 5, 10, 15 or 30 minutes. Another set of cells were incubated under hypoxia for 1 h and then returned to normoxia for 5, 10, 15 or 30 min (re-oxygenation). A control under normoxic (N) conditions was also included. The same amount of protein was analyzed by western blotting as previously described with the addition of GAB1 and its phosphorylated form. (C) MCF10A cells were placed under normoxic or hypoxic conditions for 1 h or hypoxia for 1 h and then normoxia for 10 minutes (reoxygenation). Cells were then treated at the indicated time with 10 ng/mL of HGF/SF. Cell lysates were incubated for AlphaScreen specific phospho-Erk and phospho-Akt quantitation. Error bars represent standard deviations (n = 3; ± SD). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29930749), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Downregulation of La autoantigen, hVAP-A, PSMA7, and NS5B leads to reduction in viral replication: overnight grown Huh-7.5 cells were transfected with La autoantigen, hVAP-A, and PSMA7 siRNAs for 48 h. Cells were then infected with the virus and again transfected with same siRNAs. After incubation total RNA was isolated and subjected to RT-PCR. NS5B siRNA was transfected once only after cells were infected. (a), (c), (e), and (g) represent RT-PCR results for target genes and NS5B gene upon treatment with La autoantigen, hVAP-A, PSMA7, and NS5B siRNAs, respectively. Densitometric analysis of (a), (c), (e), and (g) images is expressed in percentage in (b), (d), (f), and (h), respectively. The data represent mean ± standard deviation. ∗P value < 0.05 versus control considered statistically significant. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/27446609), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

FUS does not bind VAPB or PTPIP51 in immunoprecipitation assays from transfected HEK293 cellsCells were transfected as indicated with control vector (CTRL), HA‐FUS + CTRL, myc‐VAPB + CTRL, or myc‐VAPB + either HA‐FUS, HA‐FUSR521C or HA‐FUSR518K. VAPB was immunoprecipitated via the myc‐tag and the samples probed on immunoblots for VAPB using rabbit VAPB antibody and for co‐immunoprecipitating FUS via the HA tag. Input VAPB and FUS were detected using myc and HA antibodies.Cells were transfected as indicated with control vector (CTRL), HA‐FUS + CTRL, HA‐PTPIP51 + CTRL or HA‐PTPIP51 + either HA‐FUS, HA‐FUSR521C or HA‐FUSR518K. PTPIP51 was immunoprecipitated using rat anti‐PTPIP51 and the samples probed for PTPIP51 using rabbit anti‐HA antibody and for co‐immunoprecipitating FUS using rabbit FUS antibody. Input PTPIP51 and FUS were detected using PTPIP51 and EGFP antibodies, respectively. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/27418313), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

APCDD1 is co-regulated by MEIS1 and EWS-FLI1 and mediates the oncogenic role of MEIS1. (A, B) Silencing of (A) MEIS1 or (B) EWS-FLI1 downregulated expression of both APCDD1 mRNA and protein. Error bars represent mean ± SD of three replicates (**P < 0.01, ***P < 0.001). (C, D) APCDD1 knockdown potently (C) inhibited cell proliferation and (D) decreased colony formation. Error bars represent mean ± SD of three replicates (*P < 0.05, **P < 0.01, ***P < 0.001). (E) A673 and SKNMC cells stably expressing shMEIS1 were transiently transfected with plasmid encoding APCDD1, and subjected to immunoblotting and MTT assays. Bars represent mean ± SD of three replicates (**P < 0.01). (F) Proposed model showing that MEIS1 and EWS-FLI1 co-operatively activate APCDD1 transcription, thereby promoting the malignant phenotype of Ewing sarcoma cells. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30496486), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Pharmacologic inhibition of ATM rescues oxidative stress-induced senescence by suppressing ATM- and NEMO-mediated NF-kappa B activation. (A) Representative images of primary WT and Ercc1-/- MEFs were induced to undergo senescence by serial passaging at 20% oxygen. At passage 5, MEFs were grown in the presence or absence of KU-55933 (10 μM) for 72 hrs. Senescence was determined by SA-beta gal staining. Images were obtained at the magnification of 10x. (B) Quantitation of the percent SA-beta gal positive cells. Graph represents the mean +/- s.e.m. of three independent experiments. Student’s t-test, ***p <0.001, ****p <0.0001. (C) Passage 5 Ercc1-/- MEFs treated with vehicle or KU-55933 (10 μM) for 72 hours were collected and levels of p21 and p16INK4a were determined by western blotting. (D) Passage 5 Ercc1-/- MEFs were treated with KU-55933 (10 μM) for 72 hours and whole cell lysate (CL) and nuclear extracts (NE) were analyzed by immunoblotting for expression of proteins involved in the DNA damage response. (E) Whole cell lysate (CL) and nuclear extract (NE) were extracted from Ercc1-/- MEFs treated with 10 μM of KU-55933 for analysis of nuclear NEMO and p65. GAPDH was used as a loading control of total proteins and LaminA/C as a loading control of nuclear protein. (F) Passage 5 WT and Ercc1-/- MEFs transfected with a NF-kappa B-luciferase reporter construct were cultured in the presence or absence of KU-55933 (10 μM) and were collected for luciferase assays after 72 hours. (G) qRT-PCR analysis of mRNA expression in passage 5 WT and Ercc1-/- MEFs treated with or without of KU-55933 (10 μM) for 72 hrs. P values were determined using a Student’s t-test. *p<0.05, **p<0.01, ***p <0.001. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32201398), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Inhibition of splicing downregulates repair factors at both the mRNA and protein levels. (a) U2OS cells were treated with pladienolide B for 2, 6 and 16 h and irradiated (6 Gy, 1 h recovery) 1 h prior to termination of the treatment. The levels of the indicated mRNAs were measured through qPCR analysis. The change is relative to the DMSO control and two reference genes (18S rRNA and beta -actin). Means±S.D. are shown, n=3. (b) qPCR analysis of the levels mRNA and pre-mRNA for RNF8 and RAD51 in U2OS cells treated with pladienolide B or isoginkgetin for 2, 6 or 16 h and irradiated (6 Gy, 1 h recovery) 1 h prior to termination of this treatment. The change is expressed relative to the DMSO control value and the levels of mRNA for two reference genes (18S rRNA and beta -actin). Means±S.D. are shown, n=3. The arrows indicate the positioning of the PCR primers used and their sequences are shown in Supplementary Table S1. (c) Western blotting following pladienolide B treatment of U2OS cells as described in (a). beta -Actin was used as a loading control. Densitometric quantification of each protein is shown in Supplementary Figure S2B. The three MDC1 bands correspond to the unphosphorylated, phosphorylated and hyperphosphorylated forms of full-length MDC1 Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/27315300), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

LAIR-1 inhibits Glut1-related glucose uptake in OS cells. a Heatmap showing the levels of differentially expressed mRNAs. b Top 20 KEGG pathway annotation categories for target gene functions of predicted mRNAs. c Selected significantly differentially expressed mRNA-related to EMT in RNA-seq data between two groups, ***P < 0.001. d qPCR validation of differentially expressed EMT-related genes in LV-NC and LV-LAIR-1-overexpressing OS cells, **P < 0.01. e Glut1 expression analyzed by western blotting. f Immunofluorescence staining of Glut1 in the LV-LAIR-1-overexpressing OS cells. Scale bar = 50 μm. Data were obtained from at least two independent experiments. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32563267), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Lysyl Oxidase (LOX) expression in malignant and non-malignant human prostate tissue.(A) Representative immunohistochemical staining of LOX (brown) in sections of non-malignant prostate tissue specimens (TINT epithelium and TINT stroma) from two prostate cancer patients (original magnifications x 200). Case A show weak epithelial staining (score 1) and Case B strong epithelial staining (score 3). (B) Consecutive sections from non-malignant prostate tissue stained with in situ hybridization or immunohistochemistry for LOX. Note that mRNA (brown dots) and protein (brown) expression in individual glands were related, in the stroma in contrast mRNA was generally low but protein often detected (original magnifications x200). (C) LOX mRNA expression in non-malignant prostate tissue (TINT) (n = 12), in primary prostate tumor tissue (n = 12), and in castration-resistant bone metastases (CRPC) (n = 30) using Illumina gene expression array. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/26501565), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Recruitment of 53BP1 and TRF2 to fs NIR laser-induced DNA lesions. (A) EGFP-53BP11220-1711 and TRF2-YFP cell lines were subjected to NIR laser microirradiation with the input power ranging from 20 mW to 155 mW (corresponding to ≈7 × 1010 W/cm2 to 5.41 × 1011 W/cm2 peak irradiance) as indicated. EGFP-53BP11220-1711 and TRF2-YFP recruitment was monitored for 15 min and 6 min p.i., respectively. The number of the cells with the recruitment and the number of cells examined are shown at the top of each histogram. Detailed microirradiation energy and irradiance calculations versus the laser input power (the power of the laser beam entering the microscope) are shown in Table 2. (B) Time course fluorescence measurement of the damage-site recruitment of EGFP-53BP11220-1711 and TRF2-YFP. PtK2 cells stably expressing EGFP-53BP11220-1711 or TRF2-YFP were damaged with either low (60 mW input power with peak irradiance of ≈2.1 × 1011 W/cm2; upper panel) or high (100 mW input power with peak irradiance of 3.49 × 1011 W/cm2; lower panel) dose of NIR laser microirradiation as indicated. For EGFP-53BP11220-1711, four and three cells were followed for 30 min for 60 mW and 100 mW damage, respectively. For TRF2-YFP, four cells each were followed for 4 min. The fluorescent intensity at the damage site was divided by that in the nucleoplasm and was subtracted by one for normalization. (C) Similar experiments as in (B) were performed following EGFP-53BP11220-1711 recruitment, and live fluorescent images of one representative cell each at indicated p.i. time points are shown. Time 0:00 refers to pre-laser condition. The irradiation site is indicated by a black line at time 0:00 and white arrowheads. Consistent results were obtained with 12 and 8 cells examined for the low (60 mW) and high (100 mW) irradiation condition, respectively. Scale bar = 10 μm. (D) Similar experiments as in (C) for TRF2-YFP. Consistent results were obtained with 11 and 12 cells examined for the low (60 mW) and high (100 mW) irradiation condition, respectively. Scale bar = 10 μm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/26424850), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Bt, Bv, Ca, Fi, Gt, Pm, Po, Pm, Fe, Pm-Or, Rb, Sh

Species Glossary

Reactivity: Hu, Mu, Rt, Bt, Bv, Ca, Fi, Gt, Pm, Po, Pm, Fe, Pm-Or, Rb, Sh

Applications:

, , , , , , , , , ,

Applications: , , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-304AF350
Alexa Fluor 405	NB100-304AF405
Alexa Fluor 488	NB100-304AF488
Alexa Fluor 532	NB100-304AF532
Alexa Fluor 594	NB100-304AF594
Alexa Fluor 647	NB100-304AF647
Alexa Fluor 700	NB100-304AF700
Alexa Fluor 750	NB100-304AF750
Allophycocyanin	NB100-304APC
Allophycocyanin/Cy7	NB100-304APCCY7
Biotin	NB100-304B
DyLight 350	NB100-304UV
DyLight 405	NB100-304V
DyLight 488	NB100-304G
DyLight 550	NB100-304R
DyLight 594	NB100-304DL594
DyLight 650	NB100-304C
DyLight 680	NB100-304FR
DyLight 755	NB100-304IR
FITC	NB100-304F
HRP	NB100-304H
Janelia Fluor 549	NB100-304JF549
Janelia Fluor 646	NB100-304JF646
PE	NB100-304PE
PE/Atto594	NB100-304PEATT594
PE/Cy5.5	NB100-304PECY55
PE/Cy7	NB100-304PECY7
PerCP	NB100-304PCP
mFluor Violet 450 SE	NB100-304MFV450
mFluor Violet 500 SE	NB100-304MFV500
mFluor Violet 610 SE	NB100-304MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-304SS

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H00008878-M01

p62/SQSTM1 Antibody (2C11)

(9 Reviews)

(373 Publications)

0.1 mg

H00008878-M01

Immunocytochemistry/Immunofluorescence: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - ICC-IF analysis of SQSTM1 protein in HeLa cell using SQSTM1/p62 antibody (clone 2C11) at 10 ug/ml concentration.

Immunocytochemistry/Immunofluorescence: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Analysis of p62 in SH-SY5Y cells using anti-p62/SQSTM1 antibody. Red - p62 puncta; Blue - nuclear DAPI; Green - Cytoskeleton. Image from verified customer review.

Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Disruption of the VANGL2-p62/SQSTM1 interaction in breast cancer cells. Soft agar colony formation of T47D cells overexpressing GFP, GFP-VANGL2 and GFP-p62/SQSTM1 (right). Protein expression was revealed with anti-p62/SQSTM1, anti-VANGL2 and anti-GFP antibodies by western blot analysis (right). In anti-GFP blot, the arrowhead indicates position of co-migrating GFP-VANGL2 and GFP-p62/SQSTM1 and the asterisk pinpoints GFP alone. Error bars represent mean+/-s.d. (n=3). Image collected and cropped by CiteAb from the following publication (//www.nature.com/doifinder/10.1038/ncomms10318), licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Analysis of p62/SQSTM1 antibody on HeLa cells. Image from verified customer review.

Western Blot: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Confirmation of loss of hepatic ATG7 protein and a phenotype of hepatic autophagy inactivation. Fourth ug of liver protein were loaded on 4-20% gradient Tris-Glycine gels and subsequently immunoblotted with antibodies against ATG7, p62 and LC3. Blots were visualized with G:BOX Chemi XRQ (SynGene). Representative immunoblots are shown for WT and KO mice. Full-length immunoblots are presented in Supplementary Figure S1. ATG7: autophagy-related protein 7; p62: sequestosome 1; LC3: microtubule-associated protein 1 light chain 3; WT: Mx1-Cre-Atg7F/F mouse; KO: Mx1-Cre+Atg7F/F mouse. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-14405-w), licensed under a CC-BY license.

ELISA: p62/SQSTM1 Antibody (2C11) [H00008878-M01] - Detection limit for recombinant GST tagged SQSTM1 is approximately 0.03ng/ml as a capture antibody.

Confirmation of loss of hepatic ATG7 protein and a phenotype of hepatic autophagy inactivation. Fourty ug of liver protein were loaded on 4-20% gradient Tris-Glycine gels and subsequently immunoblotted with antibodies against ATG7, p62 and LC3. Blots were visualised with G:BOX Chemi XRQ (SynGene). Representative immunoblots are shown for WT and KO mice. Full-length immunoblots are presented in Supplementary Figure S1. ATG7: autophagy-related protein 7; p62: sequestosome 1; LC3: microtubule-associated protein 1 light chain 3; WT: Mx1-Cre-Atg7F/F mouse; KO: Mx1-Cre+Atg7F/F mouse. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29074879), licensed under a CC-BY licence.

miR-29a downregulates TFEB and ATG9A to inhibit autophagy. D). 5ug of total protein cell lysates from Panc-1 cells were transfected with CTRL, miR-29a mimics, siCTRL, siTFEB, or siATG9A. 24 hours post-transfection, total protein was harvested and subjected to western blot analysis for p62 and LC3B, and GAPDH was used as loading control. Quantification of band intensities normalized to GAPDH and relative to control are shown below respective blots. All experiments were repeated 3 times and representative data is presented. Image collected and cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.11928), licensed under a CC-BY licence.

$Time-dependent effects of subacute administration of effective dose of escitalopram (5 mg/kg/day) for 3 and 7 days on expression of 5-HT7R in the thalamic plasma membrane fraction (Panel (A)). Ordinate: mean ± SD (n = 6) of the relative protein level of 5-HT7R in the thalamic plasma membrane fraction. Panel (B) indicates the pseudo-gel images using capillary immunoblotting. ** p < 0.01 vs. control by one-way analysis of variance with Tukey’s post-hoc test. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/33572981), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

TAZ stimulates exercise-induced mitochondrial biogenesis and increases respiratory metabolism and activity.a Wild-type (WT) and muscle-specific TAZ knockout (mKO) mice were subjected to endurance exercise training. The gastrocnemius muscle obtained from these mice was isolated and embedded in paraffin. The sectioned tissues were immunostained with an anti-COX4 antibody to visualise the mitochondria. Scale bar = 50 μm. b Genomic DNA was isolated from the gastrocnemius muscle of mice described in (a). Relative mitochondrial DNA copy number was determined via quantitative PCR using primers for mitochondrial-encoded Cox2 and nuclear-encoded beta -globin. The Ct values of mitochondrial Cox2 were normalized to those of nuclear beta -globin; n = 6 for each genotype (WT untrained vs. mKO untrained; *p = 0.0188, WT untrained vs. WT trained; **p = 0.0069, WT trained vs. mKO trained; ***p = 0.0002). c Protein from the gastrocnemius muscle of mice described in (a) was assessed via immunoblotting to observe the indicated proteins. GAPDH was used as the loading control. d RNA was isolated from the gastrocnemius muscle of mice described in (a). Mitochondrial-encoded marker gene expression was analysed via quantitative reverse transcription (qRT)-PCR, n = 6 for each genotype (mt-Atp6; **p = 0.0035 for WT untrained vs. mKO untrained, **p = 0.0071 for WT untrained vs. WT trained, ****p < 0.0001 for WT trained vs. mKO trained, mt-Co2; *p = 0.0174 for WT untrained vs. mKO untrained, ****p < 0.0001 for WT untrained vs. WT trained, ****p < 0.0001 for WT trained vs. mKO trained, mt-Cytb; *p = 0.0214 for WT untrained vs. mKO untrained, *p = 0.0124 for WT untrained vs. WT trained, ****p < 0.0001 for WT trained vs. mKO trained, mt-Nd5; *p = 0.0186 for WT untrained vs. mKO untrained, **p = 0.0059 for WT untrained vs. WT trained, ****p < 0.0001 for WT trained vs. mKO trained). e Nuclear-encoded mitochondrial gene expression in the gastrocnemius muscle of mice in (a) was analysed via qRT-PCR, n = 6 for each genotype (Atp5a1; *p = 0.0137 for WT untrained vs. WT trained, **p = 0.0048 for eKO untrained vs. eKO trained, Idh3g; **p = 0.0089 for WT untrained vs. WT trained, Ndufa10; *p = 0.0205 for WT untrained vs. WT trained, Uqcrc1; ****p < 0.0001 for WT untrained vs. WT trained, **p = 0.0011 for mKO untrained vs. mKO trained). f The respiratory metabolism of mice described in (a) was analysed using the Oxylet system. Data are shown as the average values observed under light and dark conditions; n = 6 for each condition (VO2-light; **p = 0.0033 for WT untrained vs. mKO untrained, **p = 0.0098 for WT trained vs. mKO trained, VO2-dark; *p = 0.0146 for WT untrained vs. mKO untrained, ***p < 0.0007 for WT untrained vs. WT trained, ****p < 0.0001 for WT trained vs. mKO trained, VCO2-dark; ****p < 0.0001 for WT untrained vs. WT trained, ****p < 0.0001 for WT trained vs. mKO trained, Energy expenditure-light; *p = 0.0234 for mKO untrained vs. mKO trained, Energy expenditure-dark; *p = 0.021 for WT untrained vs. mKO untrained, ***p = 0.0001 for WT untrained vs. WT trained, ****p < 0.0001 for WT trained vs. mKO trained). Ten to twelve-week-old mice were used for all panels. Data are presented as mean ± SEM for (b, d–f). Statistical significance was analysed via two-way ANOVA with Sidak’s multiple comparison test. Representative data was shown and experiments were performed at least twice with similar results for (a, c). Source data are provided as a Source data file. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35115527), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry analysis of histone induced activations of caspase 1, caspase 3, and NLRP3 (400×). BMECs were cultured briefly on cover glasses (pre-treated with poly-L –lysine, 0.1 mg/mL, Sigma-Aldrich) and incubated with histone (50 and 100 μg/mL) for 16 h. The samples were visualized with DAB, counterstained with hematoxylin and observed by inverted microscope. Three independent experiments were carried out by light microscope analyses. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31156617), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Bv, Ha, Rb

Species Glossary

Reactivity: Hu, Mu, Rt, Bv, Ha, Rb

Applications:

, , , ,

Applications: , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Conjugate:

Unconjugated

Validated by:

Validated by:

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NBP1-04676

PGC1 alpha Antibody - BSA Free

(11 Reviews)

(261 Publications)

0.1 ml

NBP1-04676

Mitochondrial content and biogenesis. Immunoblot analysis of PGC1 alpha in MEFs (n = 7; t = 5.764; df = 6). Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-017-12662-3), licensed under a CC-BY license.

Analysis of a FFPE section of mouse prostate using rabbit polyclonal PGC1 alpha Antibody at 1:200 dilution. The antibody generated an expected strong nuclear (punctate in some cells) and a weak cytoplasmic staining in the glandular cells lining of tubule-alveolar gland.

An intracellular stain was performed on A431 cells with PGC1 alpha Antibody NBP1-04676 (blue) and matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

<b>Biological Strategies Validation. </b>Vehicle or RU 486 (5 mg/kg) injected mice were presented with/without corticosterone (10 mg/kg) for 3 days. The expressions of peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1 alpha NBP1-04676) and NIX (BNIP3L NBP1-88558) were visualized via western blotting. Loading control is beta-actin. n = 5. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33473105/) licensed under a CC-BY license.

Total protein from human adipose and skeletal muscle tissue, HeLa and A431 cells lines was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/mL PGC1 alpha Antibody (Molecular weight: 91 KDa) in 1% non-fat milk in TBST and detected with an anti-rabbit HRP secondary antibody using chemiluminescence

Western Blot of total protein from murine skeletal muscle tissue using PGC1 alpha Antibody. Image from verified customer review.

HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with PGC1 alpha Antibody [NBP1-04676] at a 1:200 dilution overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) [NB100-690] was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse DyLight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Analysis of PGC1 alpha Antibody in mouse prostate using DAB with hematoxylin counterstain.

An intracellular stain was performed on HeLa cells with NBP1-04676AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

An intracellular stain was performed on HepG2 cells with PGC1 alpha Antibody and a matched isotype control. Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody.

Western blot shows lysates of A431 human squamous carcinoma parental cell line and PGC1 alpha knockout (KO) A431 cell line. PVDF membrane was probed with 1:1000 of Rabbit Polyclonal PGC1 alpha Antibody (Catalog # NBP1-04676) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog #HAF008). Specific band was detected for PGC1 alpha at approximately 105 kDa (as indicated) in the parental A431 cell line, but is not detectable in the knockout A431 cell line. This experiment was conducted under reducing conditions.

DM1-derived fibroblasts have no changes in mitochondria biogenesis. (A, B) Representative images of immunofluorescence of TOMM20, and PGC1-alpha in DM1 and control fibroblasts (n=3). (C) Medium fluorescence intensity of MitoTracker Red FM in control (n=3) and DM1 cells (n=5) and (D) of Rhodamine 123 in DM1 and control fibroblasts (n=3). (E) mRNA levels of TFAM transcription factor (n=3). (F) mRNA levels of OPA1, MFN1, MFN2, DRP1 and PARKIN in DM1 and control fibroblasts (n≥2).

Feeding the diabetes mellitus (DM) mice with the RSV-enriched diet improved mitochondrial biogenesis in the quadriceps muscle. A) Representative western blots using antibodies against nuclear respiratory factor (NRF-1), ATP synthase, H+ transporting, mitochondrial F1 complex, beta polypeptide (ATP5B), cytochrome c oxidase subunit IV (Cox IV), peroxisome proliferator-activated receptor-gamma coactivator 1-alpha (PGC-1alpha), and GAPDH. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29578301), licensed under a CC-BY licence.

Mitochondrial content and biogenesis. (d) Immunoblot analysis of PGC1alpha in MEFs (n = 7; t = 5.764; df = 6). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28970491), licensed under a CC-BY licence.

Atg7 deficiency in vascular smooth muscle cells (VSMCs) alters mitochondrial quality control and enhances apoptosis. c) Western blot analyses of the expression of TFEB and PGC-1-alpha proteins in aortic VSMCs isolated from Atg7+/+Tagln-Cre+, ApoE-/- and Atg7F/FTagln-Cre+, ApoE-/- mice after 10 weeks of HFD. Cells were incubated either with or without oxidized LDL (200 ug ApoB/mL) for 8 h. beta-Actin was used as the loading control. The graph represents the densitometric analysis of the expression level of TFEB and PGC-1-alpha proteins. The data are expressed as mean +/- SEM of four independent experiments from different primary VSMC cultures. *P 12 in each group. (B) Western blot images using S100 beta , tubulin beta‐3, and beta ‐actin antibodies to total protein lysate in hippocampal area. The statistical results of internal standardized S100 beta / beta ‐tubulin ratio by beta ‐actin levels relative to young adult wild‐type C57BL/6J. White and black bars indicate the wild‐type C57BL/6J and Tet‐mev‐1 mice, respectively. Data are expressed as mean ± SD; *P 12 in each group. (C) Micrographs of immunohistochemical analysis on paraffined hippocampal tissue sections using GFAP and S100 beta antibody. Brown cells indicate GFAP‐stained astrocytes. Scale bar = 100 μm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/27623715), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

At the time of second passage, vessels in successfully growing xenografts of four different tumor types surveyed were of murine origin: A-C Colon; D-F Lung; G-I Pancreatic; J-L Renal Cell Carcinoma. For each tumor type, a representative section of an original patient specimen labeled with anti-huCD31 is shown (A, D, G, J). For each tumor, sections of the first passage xenografted tumor when it was resected are also shown; while no huCD31(+) vessels were identified in the xenografts (B, E, H, K), msCD34(+) vessels were abundant (C, F, I, L). (bars = 50 μ). Image collected and cropped by CiteAb from the following open publication (//translational-medicine.biomedcentral.com/articles/10.1186/1479-5876-11-110), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Knockdown of autophagy-related gene 5 (ATG5) inhibits trehalose-induced autophagy in normal human primary airway epithelial cells.Normal human tracheobronchial epithelial cells were transfected with Naito1 chimera RNAi (control siRNA) or ATG5 chimera siRNA (ATG5 siRNA). Twenty-four hours after siRNA transfection, cells were treated with medium or trehalose (TRE, 100 mM) for 48 h. ATG5 protein (A) and LC3 I and LC3 II proteins (B) were examined by Western blot analysis with GAPDH protein used as loading control. The representative Western blot picture was shown from 2 independent experiments with each being performed in triplicate wells. Image collected and cropped by CiteAb from the following open publication (//dx.plos.org/10.1371/journal.pone.0124524), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$Expression of ER stress associated protein and autophagy related protein in the C28I2 chondrocyte infected with Ad-ATG5 or Ad-ATG7. a The ATG5 and ATG7 mRNA level were analysed by RT-PCR in C28I2 cells. b Analysis of ATG5 and ATG7 mRNA level with Q-PCR in C28I2 cells. Ad-ATG5(a), Ad-ATG7(b). c The ATG5 protein expression was analysed by western blotting in the Ad-ATG5 infected C28I2 cells. The level of ATG5 was normalized to beta -actin. d The ATG7 protein expression was analysed by western blotting in the Ad-ATG7 infected C28I2 cells. The level of ATG7 was normalized to beta -actin. e The expression levels of LC3, P62, PERK, p-PERK, Nrf2 proteins after infected with Ad-ATG5 in C28I2 cells for 24 h by western blotting. The levels of related proteins were normalized to beta -actin. f The expression levels of LC3, P62, PERK, p-PERK, Nrf2 proteins after infected with Ad-ATG7 in C28I2 cells for 24 h by western blotting. The levels of related proteins were normalized to beta -actin. *P 100 mitochondria). Data analysis was performed by using one-way ANOVA (∗P 250 cells/chains per sample). c Representative whole-mount IF of adult Plzf-mC/CreER; Z/EG tubules D3 post-TAM (n = 2 mice). Arrowheads: PDX1+ As. Inset shows detail of indicated area. Scale bar, 50 μm. d Representative IF of adult WT testis sections (n = 4 mice). Insets: details of selected areas. EOMES+ GFR alpha 1+ (arrowheads) and EOMES− GFR alpha 1+ cells (bracket) are shown. Tubule stage is indicated. Scale bar, 50 μm. e Representative IF of adult Oct4-GFP testis section (n = 2 mice). Insets: immunostaining within indicated area. EOMES+ GFP− (arrowheads) and EOMES− GFP+ (bracket) spermatogonia are indicated. Asterisks: autofluorescent interstitium. Scale bar, 50 μm. f Representative flow cytometry of fixed and permeabilized testis cells from Oct4-GFP adults (n = 2). PLZF+ cells shown. g Representative IF of adult WT testis sections (n = 4 mice). Insets: immunostaining within indicated area. PDX1+ EOMES+ spermatogonia are indicated (arrowheads). Tubule stage is shown. Asterisk: autofluorescent interstitium. Scale bar, 50 μm. h Representative flow cytometry of fixed and permeabilized WT adult testis cells (n = 4). Mean numbers of EOMES+ and PDX1+ cells in PDX1+ and EOMES+ gates respectively are shown ± s.e.m. i Representative flow cytometry for KI67 in indicated PLZF+ populations of fixed and permeabilized adult WT testis cells (n = 4 mice). j Quantification of flow cytometry from i. Percentage of indicated PLZF+ fractions KI67+ . Horizontal bars: mean values (n = 4 mice). k Representative IF of adult WT testis sections demonstrating PDX1+ Aundiff localisation (arrowheads) within tubules (n = 4 mice). Asterisks: autofluorescent interstitium. Tubule stages are shown. Scale bar, 50 μm. l Quantification of spermatogonial localisation from k. Mean values ± s.e.m. are shown (n = 4 mice, 56–95 tubule sections per mouse). Significance vs. tubule-tubule localisation is indicated. m Representative IF of Id4IRES-GFP adult testis sections (n = 4 mice). Insets show immunostaining within indicated area. Arrowheads: PDX1+ EOMES+ GFP+ spermatogonia. Asterisk: PDX1low EOMES+ GFP+ cell. Tubule stage is shown. Scale bar, 50 μm. n Representative flow cytometry of indicated PLZF+ populations of fixed and permeabilized adult Id4IRES-GFP testis cells (n = 4 mice). Mean numbers of PDX1+ and EOMES+ Aundiff expressing GFP ± s.e.m. o Flow cytometry of Id4IRES-GFP testis cells as in n. PLZF+ fractions are shown. Mean numbers of PLZF+ GFP+ cells positive for EOMES and PDX1 are indicated ± s.e.m. (n = 4 mice). Significance was calculated by two-tailed Student’s t-test (***P < 0.001, ****P < 0.0001) Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/s41467-018-04827-z), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Reactivity:

Hu, Mu, Rt, Po, Gt, Ha, Sh, Sq, Ca, Eq, Pm

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Gt, Ha, Sh, Sq, Ca, Eq, Pm

Applications:

, , , , , , , ,

Applications: , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP1-04676AF350
Alexa Fluor 405	NBP1-04676AF405
Alexa Fluor 488	NBP1-04676AF488
Alexa Fluor 532	NBP1-04676AF532
Alexa Fluor 594	NBP1-04676AF594
Alexa Fluor 647	NBP1-04676AF647
Alexa Fluor 700	NBP1-04676AF700
Alexa Fluor 750	NBP1-04676AF750
Allophycocyanin	NBP1-04676APC
Biotin	NBP1-04676B
DyLight 350	NBP1-04676UV
DyLight 405	NBP1-04676V
DyLight 488	NBP1-04676G
DyLight 550	NBP1-04676R
DyLight 594	NBP1-04676DL594
DyLight 650	NBP1-04676C
DyLight 680	NBP1-04676FR
DyLight 755	NBP1-04676IR
FITC	NBP1-04676F
HRP	NBP1-04676H
Janelia Fluor 549	NBP1-04676JF549
Janelia Fluor 646	NBP1-04676JF646
PE	NBP1-04676PE
PerCP	NBP1-04676PCP
mFluor Violet 450 SE	NBP1-04676MFV450
mFluor Violet 500 SE	NBP1-04676MFV500
mFluor Violet 610 SE	NBP1-04676MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NBP1-04676SS

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NB100-182

FANCD2 Antibody - BSA Free

(8 Reviews)

(215 Publications)

0.05 ml

NB100-182

<b>Biological Strategies Validation. </b>Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - AMPK-activating AICAR treatment activates FANCD2, a pivotal molecule of Fanconi anemia DNA damage signaling pathway. AICAR treatment induces FANCD2 monoubiquitination in transformed normal fibroblasts (GM00637I). GM00637I cells were treated with 1 mm 2-deoxyglucose, 0.25 mM AICAR, or 1 mm phenformin for 24 h. Lysates were subjected to western blotting with FANCD2 Antibody, phospho-AMPKalpha1 (T172), and AMPKalpha and beta-actin. In FANCD2 blots, the position of monoubiquitinated FANCD2 (Ub-FANCD2) is indicated by an arrow. Image collected and cropped by CiteAb from the following publication (//doi.wiley.com/10.1002/2211-5463.12185) licensed under a CC-BY license.

Simple Western: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - Simple Western lane view shows a specific band for FANCD2 in 0.1 mg/ml of HeLa lysate using FANCD2 Antibody. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Immunocytochemistry/Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - Analysis using the Biotin conjugate of FANCD2 Antibody. FANCD2 colocalizes in vivo with another protein in SiHa cells after cell exposure to IR. Proliferating SiHa cells were exposed to 10 Gy of IR and double color immunofluorescence staining was performed after 8 h. Images were captured in a Kodak digital image system on a Leica fluorescence microscope.

<b>Biological Strategies Validation. </b>Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - AICAR-induced FANCD2 monoubiquitination is dependent on AMPK. Inhibitor of AMPK blocks AICAR-induced FANCD2 monoubiquitination in GM00637I normal fibroblasts. Cells were pretreated with 5 um of Compound C (an AMPK inhibitor) 1 h before treatment with 0.25 mm AICAR for 24 h. Cell lysates were subjected to immunoblotting with FANCD2 Antibody to visualize monoubiquitinated FANCD2 (Ub-ANCD2). Image collected and cropped by CiteAb from the following publication (//doi.wiley.com/10.1002/2211-5463.12185) licensed under a CC-BY license.

Western Blot: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - The FA-BRCA Pathway Is Activated on the Sex Chromosomes during Meiosis. Schematic of the FA-BRCA pathway. FA proteins analyzed in this study are shown in color. Western blot analysis with three independent anti-FANCD2 antibodies (G33, E33, and Novus NB100-182 antibody: NB). K561R, PD20 cells expressing a mutated form of FANCD2 incapable of monoubiquitination; WT, PD20 cells complemented with wild-type FANCD2; Vector, PD20 cells containing empty vector. Image collected and cropped by CiteAb from the following publication (//linkinghub.elsevier.com/retrieve/pii/S2211124716313298), licensed under a CC-BY license.

<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - HES1K109E mutation does not impair MMC-induced HES1 foci formation. HeLa cells transfected with HA-tagged HES1 or HES1K109E coding vectors were processed for immunofluorescence 16 h following treatment with MMC (120 ng/ml). Cells were double-stained with anti-HA and anti-FANCD2 antibodies (mouse anti-HA at 1:500; rabbit anti-FANCD2, Novus Biologicals NB100-182 at 1:2000) followed with secondary antibodies (Goat anti-mouse Alexafluor-555, ThermoScientific A-32727 and goat anti-rabbit Alexafluor-488, ThermoScientific, A-11008). Cell nuclei were labelled with TO-PRO-3 Iodide stain (1:400, ThermoScientific, T3605). Cells were visualized at 100x magnification. Image collected and cropped by CiteAb from the following publication (//bmcresnotes.biomedcentral.com/articles/10.1186/s13104-018-3243-7), licensed under a CC-BY license.

Immunohistochemistry: FANCD2 Antibody - BSA Free [NB100-182] - FANCD2 Antibody [NB100-182] - The tissue sections were incubated with a primary antibody cocktail of FANCD2 Antibody (NB100-182) at a dilution of 1:1000 & a monoclonal anti-Ki67 mouse antibody at a dilution of 1:150 for 1 h at room temperature. Then the slides were incubated with a secondary antibody cocktail containing FITC conjugated anti-rabbit IgG and Alexafluor 594 donkey anti-mouse secondary. The sections were mounted on glass slides using a DAPI-containing embedding medium (Vysis Dapi 1, Abbott Laboratories, Downers Grove, IL, USA), then analyzed under a fluorescence microscope. FANCD2 (top) foci positive NSCL tumor, and FANCD2 (bottom)foci negative NSCL tumor. Magnification: 1000x. Image collected and cropped by CiteAb from the following publication (//journal.frontiersin.org/article/10.3389/fonc.2014.00368/abstract), licensed under a CC-BY license.

Western Blot: FANCD2 Antibody [NB100-182] - Analysis of FANCD2 (Molecular weight: 164.1 KDa) using the HRP conjugate of FANCD2 Antibody (lot C) in HeLa WCE.

Western Blot: FANCD2 Antibody [NB100-182] - Functional assessment of Fanconi anemia pathway. Western blot with FANCD2 Antibody of non-FA control lymphoblasts (LCL), proband LCL-A-2017, proband LCL-B-2017, FANCB mutant (null) LCL, and FANCD2 mutant (null) LCL. Image collected and cropped by CiteAb from the following publication (//doi.wiley.com/10.1002/mgg3.350), licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: FANCD2 Antibody [NB100-182] - Mouse Embryonic Fibroblasts Untreated and treated with Mitomycin C for 24hr with FANCD2 Antibody diluted 1:500 in 15%BCS+0.3% Triton X 100 in PBS. This image was submitted via customer Review.

Immunohistochemistry: FANCD2 Antibody [NB100-182] - Staining of human prostate, glandular epithelium using FANCD2 Antibody. Image using the Biotin format of this antibody.

Representative images of the cellular response to RS (induced by a low dose of hydroxyurea; HU) in U2OS cells, as determined by IF staining for gamma H2AX and FANCD2A selected cell is defined by the yellow arrow, and is enlarged in the bottom panel. Scale bars are indicated.

Chemical library screening for small molecules that inhibit the Fanconi anemia pathway. (A) Schematic of the screening for small molecules that inhibit IR-induced FANCD2 foci formation. (B) Representative photomicrographs of EGFP-FANCD2 foci in PD20F-EGFP-FANCD2 cells untreated and treated with the indicated compounds at the indicated concentration. The cells were treated with compounds immediately before irradiation (15 Gy), and fixed after 12 hours. (C) FANCD2 foci in HeLa cells untreated and treated with the indicated compounds at the indicated concentration. The cells were fixed 8 hours after irradiation (10 Gy) and immunostained with anti-FANCD2 antibody. Scale bar = 20 um. Image collected and cropped by CiteAb from the following publication (//molecular-cancer.biomedcentral.com/articles/10.1186/1476-4598-11-26), licensed under a CC-BY licence.

The FA-BRCA Pathway Is Activated on the Sex Chromosomes during Meiosis. (F) Western blot analysis with three independent anti-FANCD2 antibodies (G33, E33, and Novus NB100-182 antibody: NB). K561R, PD20 cells expressing a mutated form of FANCD2 incapable of monoubiquitination; WT, PD20 cells complemented with wild-type FANCD2; Vector, PD20 cells containing empty vector. Image collected and cropped by CiteAb from the following publication (//linkinghub.elsevier.com/retrieve/pii/S2211124716313298), licensed under a CC-BY licence.

HES1 mono-ubiquitination is not required for foci formation. a HES1K109E mutation does not impair MMC-induced HES1 foci formation. HeLa cells transfected with HA-tagged HES1 or HES1K109E coding vectors were processed for immunofluorescence 16 h following treatment with MMC (120 ng/ml). Cells were double-stained with anti-HA and anti-FANCD2 antibodies (mouse anti-HA at 1:500; rabbit anti-FANCD2, Novus Biologicals NB100-182 at 1:2000) followed with secondary antibodies (Goat anti-mouse Alexafluor-555, ThermoScientific A-32727 and goat anti-rabbit Alexafluor-488, ThermoScientific, A-11008). Cell nuclei were labelled with TO-PRO-3 Iodide stain (1:400, ThermoScientific, T3605). Cells were visualized at 100x magnification. b HES1 lysine 109 plays a role in MMC-induced activation of the HES1 promoter. COS-1 cells transfected with pHES1pro-Luc and HES1 or HES1K109E coding vectors (0.034 ug) were treated with 120 ng/ml MMC. Empty vectors were used as controls. Experiments were done twice in triplicates. 2-way ANOVA with Bonferroni correction test, *p 10 larvae/sample). The blots were probed with antibodies against Lc3 and Actin as a loading control and Lc3-II/Lc3-I ratios are indicated below. Western blots were repeated twice with independent extracts. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30818338), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Ddx19 is specifically required for the nuclear localization of MKL1.(a) Western blotting (WB) of cells treated with control (Ctrl) or Ddx19 siRNAs (si). (b) Localization of MKL1-GFP (left panels) and endogenous MKL1 ( alpha -MKL1; right panel) in serum (FBS)-stimulated NIH 3T3 cells. Scale bars, 20 μm. (c) Quantification of MKL1-GFP localization in serum-starved (0.3% FBS), serum-stimulated (15% FBS) and LMB-treated control (−) or Ddx19 (+) siRNA-transfected cells. C, cytoplasmic; N/C, pancellular; N, nuclear. One hundred cells per point; n=3; error bars are s.e.m. (d) Localization of nuclear proteins in NIH 3T3 cells transfected with control (−) or Ddx19 (+) siRNA. SRF (serum response factor), RanBPI-delta NES (Ran binding protein 1 lacking nuclear export signal), hnRNPU (heterogeneous nuclear ribonucleoprotein U), Smad4+LMB (Smad4 stimulated with leptomycin B), MC (myocardin). More than fifty cells per point; n=2; error bars are s.d. (e) Localization of NLS-2GFP-NES in serum-starved (0.3% FBS) or LMB-treated cells transfected with control (−) or Ddx19 (+) siRNAs. One hundred cells per point; n=2; error bars are s.d. Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/ncomms6978), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Av, Ca, Ma, Pm, Ze

Species Glossary

Reactivity: Hu, Mu, Rt, Av, Ca, Ma, Pm, Ze

Applications:

, , , , , , , , , , ,

Applications: , , , , , , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 405	NB100-182AF405
Alexa Fluor 488	NB100-182AF488
Alexa Fluor 647	NB100-182AF647
Alexa Fluor 700	NB100-182AF700
Biotin	NB100-182B
DyLight 405	NB100-182V
DyLight 488	NB100-182G
DyLight 550	NB100-182R
DyLight 650	NB100-182C
FITC	NB100-182F
HRP	NB100-182H

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-182SS

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H00023435-M01

TDP-43/TARDBP Antibody (2E2-D3)

(3 Reviews)

(210 Publications)

0.1 mg

H00023435-M01

Western Blot: TDP-43/TARDBP Antibody (2E2-D3) [H00023435-M01] - PDI co-precipitates with TDP-43/TARDBP and is increased in mutant TDP-43 transgenic mouse spinal cords. Co-immunoprecipitation of wildtype and mutant Q331K TDP-43-mCherry from Neuro2a cell lysates using an anti-PDI antibody followed by TDP-43 immunoblot. Control immunoprecipitations were performed using untransfected (Untrans) or mCherry expressing cell lysates, buffer only with coprecipitating antibodies or TDP-43 Q331K cell lysates using an irrelevant, isotype-matched control antibody (IgG). Input control (2%) shows expression of mCherry-TDP43 in wildtype and Q331K cell lysates. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0081170) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: TDP-43/TARDBP Antibody (2E2-D3) [H00023435-M01] - Analysis of monoclonal antibody to TARDBP on HeLa cell. Antibody concentration 10 ug/ml.

Immunohistochemistry-Paraffin: TDP-43/TARDBP Antibody (2E2-D3) [H00023435-M01] - TARDBP on formalin-fixed paraffin-embedded human leiomyosarcoma. [antibody concentration 3 ug/ml]

Western Blot: TDP-43/TARDBP Antibody (2E2-D3) [H00023435-M01] - Analysis of TARDBP expression in transfected 293T cell line by TARDBP monoclonal antibody (M01), clone 2E2-D3.Lane 1: TARDBP transfected lysate(44.7 KDa).Lane 2: Non-transfected lysate.

Immunohistochemistry-Paraffin: TDP-43/TARDBP Antibody (2E2-D3) [H00023435-M01] - Analysis of monoclonal antibody to TARDBP on formalin-fixed paraffin-embedded human leiomyosarcoma tissue Antibody concentration 3 ug/ml.

<b>Genetic Strategies Validation. </b>Knockdown Validated: TDP-43/TARDBP Antibody (2E2-D3) [H00023435-M01] - Analysis of TARDBP over-expressed 293 cell line, cotransfected with TARDBP Validated Chimera RNAi ( Cat # H00023435-R01V ) (Lane 2) or non-transfected control (Lane 1). Blot probed with TARDBP monoclonal antibody (M01), clone 2E2-D3 (Cat # H00023435-M01 ). GAPDH ( 36.1 kDa ) used as specificity and loading control.

ELISA: TDP-43/TARDBP Antibody (2E2-D3) [H00023435-M01] - Detection limit for recombinant GST tagged TARDBP is 0.3 ng/ml as a capture antibody.

SINE compounds partially rescue motor deficits in an animal model of ALS/FTD. (b) Western blot demonstrating the expression of human TDP43 within the brains of AAV9-injected animals, which we have previously shown is largely restricted to neuronal cell types (Jackson et al.59). Full-length blots appear in Fig. S1. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-018-22858-w), licensed under a CC-BY licence.

CLIP shows that TDP-43 directly interacts with translational target mRNAs. (C and D) Plots showing percentage of input mRNA in the TDP-43 IP for the mRNAs indicated in three separate CLIP assays from MN1 cells. Performing CLIP from UV-treated cells leads to significant recovery and enrichment relative to Gapdh for all mRNAs examined (C). In the absence of UV treatment, much less mRNA is recovered and no significant enrichment relative to Gapdh mRNA is observed. Note that the Y-axis scale in D is 1000x smaller than in C, indicating strong crosslinking dependence of mRNA co-immunoprecipitation (n = 3, s.e.m. error bars, P-values indicated in the plot, unpaired two-tailed t-test). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30357366), licensed under a CC-BY licence.

PDI co-precipitates with TDP-43 and is increased in mutant TDP-43 transgenic mouse spinal cords.(A) Co-immunoprecipitation of wildtype and mutant Q331K TDP-43-mCherry from Neuro2a cell lysates using an anti-PDI antibody followed by TDP-43 immunoblot. Control immunoprecipitations were performed using untransfected (Untrans) or mCherry expressing cell lysates, buffer only with coprecipitating antibodies or TDP-43 Q331K cell lysates using an irrelevant, isotype-matched control antibody (IgG). Input control (2%) shows expression of mCherry-TDP43 in wildtype and Q331K cell lysates. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0081170), licensed under a CC-BY licence.

Reactivity:

Hu, Mu, Rt, Ba, I, Pm, Pp

Species Glossary

Reactivity: Hu, Mu, Rt, Ba, I, Pm, Pp

Applications:

, , , , , ,

Applications: , , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Conjugate:

Unconjugated

Validated by:

Validated by:

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NBP1-48320

p62/SQSTM1 Antibody - BSA Free

(7 Reviews)

(114 Publications)

0.1 ml

NBP1-48320

<b>Biological Strategies Validation. </b>Western Blot: p62/SQSTM1 Antibody - BSA Free [NBP1-48320] - p62/SQSTM1 Antibody [NBP1-48320] - Cultured HeLa cells were treated with or without 50 uM chloriquine for 24 hours as indicated. Cell lysates were prepared and separated on a 12% gel by SDS-PAGE. Protein was transferred to PVDF membrane and blocked in 5% non-fat milk. The membrane was then probed with 1 ug/ml anti-p62/SQSMT1 in 1% milk and dectected with an anti-rabbit HRP secondary antibody using chemiluminescence. Note the upregulation of p62 (arrowhead) in response to chloroquine treatment and the blockage of autophagy.

<b>Biological Strategies Validation. </b>Western Blot: p62/SQSTM1 Antibody - BSA Free [NBP1-48320] - p62/SQSTM1 Antibody [NBP1-48320] - Immunoblotting analysis of LC3 and p62/SQSTM1 expression in thioglycollate-elicited peritoneal macrophages. Macrophages were isolated from C57BL/6 mice fed diets for 12 weeks, treated with or without 100 ng/ml LPS for 18 h with or without 50 nM bafilomycin A1. Image collected and cropped by CiteAb from the following publication (//linkinghub.elsevier.com/retrieve/pii/S0022227520339523), licensed under a CC-BY license.

<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: p62/SQSTM1 Antibody - BSA Free [NBP1-48320] - p62/SQSTM1 Antibody [NBP1-48320] - Untreated HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with anti-p628/SQSTM1 at a 1:200 dilution overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse DyLight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

<b>Biological Strategies Validation. </b>Western Blot: p62/SQSTM1 Antibody - BSA Free [NBP1-48320] - p62/SQSTM1 Antibody [NBP1-48320] - Vacuolin-1 analogues identified via virtual screening inhibited autophagic flux in HeLa cells. (A) Treatment of HeLa cells with vacuolin-1 analogues (10 M) and BAF (100 nM) failed to further increase the accumulation of both LC3B-II and SQSTM1 as compared to either drug alone. (B) Vacuolin-1 (10 M) or VS6 (10 M) induced the accumulation of yellow LC3B-II puncta in RFP-GFP-LC3B expressing HeLa cells. Scale bar = 10 m. Identification of Novel Vacuolin-1 Analogues as Autophagy Inhibitors by Virtual Drug Screening and Chemical Synthesis. Molecules (2017)

Western Blot: p62/SQSTM1 Antibody - BSA Free [NBP1-48320] - p62/SQSTM1 Antibody [NBP1-48320] - Vacuolin-1 analogues identified via virtual screening induced the accumulation of both LC3B-II and SQSTM1 in HeLa cells in a dose dependent manner after a 6 h treatment. Identification of Novel Vacuolin-1 Analogues as Autophagy Inhibitors by Virtual Drug Screening and Chemical Synthesis. Molecules (2017)

Time course of IFN signaling, autophagy, and presence of LC3 clusters in the TG after HSV-1 corneal infection. (C) (Left) p-Stat1 (Y701), Stat1, Isg15, p-Beclin-1 (T117), Beclin-1, and P62 were analyzed by WB using TG protein extracts from infected LC3-GFP+/- mice (1 x 10e6 PFU/eye, HSV-1 strain 17) during the time indicated. (Right) Quantification of WBs normalized to beta-actin. Each protein analyzed was normalized to its own beta-actin WB. n = 6 TG from 6 different mice, performed in two independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30837340), licensed under a CC-BY licence.

Autophagy signaling is induced in Vitamin C deficient neutrophils. (C) Representative Western blot for expression of p62 and actin from peritoneal PMNs of VitC sufficient and deficient Gulo-/- mice. Densitometry of normalized p62 expression from peritoneal PMNs of VitC sufficient and deficient Gulo-/- mice (N = 6 for each group, ns p = 0.3). Image collected and cropped by CiteAb from the following publication (//www.mdpi.com/2072-6643/5/8/3131), licensed under a CC-BY licence.

ROS overproduction mediated apoptosis induced by combined treatment of DDP and HIF-1 alpha in PC-3 xenografts and cell culture. (a) ROS monitored by DHE (red) and nuclei by DAPI (blue) staining in PCa xenografts (scale bars, 50 µm). (b) MDA formation of PCa xenografts was examined after various treatments (c,d). PC-3 cells were treated with DDP, si-HIF-1 alpha plasmid, or both, in the presence or absence of NAC (5 mM) or DHLA (0.25 mM) for 24 h. Total lysates (c) and culture media (d) were used to detect cellular H2O2 level. (e,f) Western analysis for HIF-1 alpha as well as cleaved caspase-3 and PARP in PC-3 cells following various treatments. Data were presented as mean ± SD of three independent experiments. *p < 0.05 versus control group; #p < 0.05 versus si-HIF-1 alpha or DDP group; $p < 0.05 versus DDP/si-HIF-1 alpha group. The original blots are presented in Supplementary Figure 8. Image collected and cropped by CiteAb from the following open publication (//www.nature.com/articles/s41598-017-07973-4), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Necroptosis marker p-MLKL can be detected in OA cartilage and after TNF/CHX stimulation.Exemplary images of impacted and differently stimulated cartilage explants after immunohistochemical staining of a cleaved CASP8, b RIPK3, and c p-MLKL. In addition, percentage of cleaved CASP8− (d) and p-MLKL− (e) positive cells was quantified (n = 3). Images were acquired using a 20× objective; the bars represent 200 µm Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31527653), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Ch, Po, Bv, Ca, ChHa, In, Sh

Species Glossary

Reactivity: Hu, Mu, Rt, Ch, Po, Bv, Ca, ChHa, In, Sh

Applications:

, , , , ,

Applications: , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP1-48320AF350
Alexa Fluor 405	NBP1-48320AF405
Alexa Fluor 488	NBP1-48320AF488
Alexa Fluor 532	NBP1-48320AF532
Alexa Fluor 594	NBP1-48320AF594
Alexa Fluor 647	NBP1-48320AF647
Alexa Fluor 700	NBP1-48320AF700
Alexa Fluor 750	NBP1-48320AF750
Allophycocyanin	NBP1-48320APC
Biotin	NBP1-48320B
DyLight 350	NBP1-48320UV
DyLight 405	NBP1-48320V
DyLight 488	NBP1-48320G
DyLight 550	NBP1-48320R
DyLight 594	NBP1-48320DL594
DyLight 650	NBP1-48320C
DyLight 680	NBP1-48320FR
DyLight 755	NBP1-48320IR
FITC	NBP1-48320F
HRP	NBP1-48320H
Janelia Fluor 549	NBP1-48320JF549
Janelia Fluor 646	NBP1-48320JF646
PE	NBP1-48320PE
PerCP	NBP1-48320PCP
mFluor Violet 450 SE	NBP1-48320MFV450
mFluor Violet 500 SE	NBP1-48320MFV500
mFluor Violet 610 SE	NBP1-48320MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NBP1-48320SS

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NB100-384

Histone H2AX [p Ser139] Antibody

(1 Review)

(156 Publications)

0.1 ml

NB100-384

Detection of Human and Mouse Histone H2AX [p Ser139] by Western Blot. Samples: Nuclear extract (50 ug) from human HEK293, human melanoma (G361), mouse wildtype embryonic fibroblasts (+/+) or mouse H2AX knockout embryonic fibroblasts (-/-). Antibody: Affinity purified rabbit Histone H2AX [p Ser139] antibody NB100-384 used at 0.1 ug/ml. Detection: Chemiluminescence with 30 second exposure. (NCS, neocarzinostatin - 200 ng/ml, 30 min). Bands appear at an observed molecular weight of ~15 kDa.

<b>Biological Strategies Validation and Genetic Strategies Validation.</b>Samples: Neocarzinostatin treated asynchronous HeLa cells (left) and untreated asynchronous HeLa cells (right) . Antibody: Affinity purified rabbit Histone H2AX [p Ser139] used at a dilution of 1:5,000 (0.2ug/ml). Detection: Red fluorescent Anti-rabbit IgG-DyLight 594 used at a dilution of 1:100.

Simple Western lane view shows a specific band for Histone H2AX [p Ser139] in 0.2 mg/ml of Jurkat lysate(s). This experiment was performed under reducing conditions using the 12 - 230 kDa separation system.

<b>Biological Strategies Validation. </b>Immunocompromised mice were subcutaneously injected with cancerous cell lines and tumors were allowed to establish. Treatments occurred every other day and the studied compound or the equivalent vehicle control administered intraperitoneally for five weeks. Tumor volume and mass were measured two times per week. IHC analysis of sectioned tumor tissues from the MDA-MB-231 study. Each section was subjected to the specified antibody followed by a biotinylated secondary antibody. Detection was done using a DAB Peroxidase HRP Substrate Kit (brown) followed by Hematoxylin counterstaining (purple). Images were obtained using inverted bright field microscopy. Sectioning results are representative of three individual tumors. Scale bar is 50 microns. Image collected and cropped by CiteAb from the following publication (nature.com/articles/s41598-017-01230-4), licensed under a CC-BY license.

Immunostaining of Histone H2AX [p Ser139] WT1 and 5mC in patients with IgA nephropathy and controls. Examples of PAS staining and immunostaining with Histone H2AX [p Ser139] (green) and WT1 (red), pATM and 5mC in glomeruli of IgA nephropathy and controls. A kidney sample of a 65-year-old male of IgA nephropathy without podocytopathic features. Arrows indicate Histone H2AX [p Ser139] and WT1 double-positive cells. Scale bars: 50um. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-019-57140-0) licensed under a CC-BY license.

<b>Biological Strategies Validation. </b>Samples: Nuclear extract from HeLa cells treated with 100 uM EPE for 4 hours (+) or mock treated (-). Antibody: Affinity purified rabbit Histone H2AX [p Ser139] antibody used at 0.1 ug/ml. Detection: Chemiluminescence with an exposure time of 3 minutes. Band appears at an observed molecular weight of ~17 kDa.

<b>Biological Strategies Validation. </b>Detection of human Histone H2AX [p Ser139] by western blot. Samples: Whole cell lysate (50 ug) from Jurkat cells treated with 100 uM EPE for 4 hours (+) or mock treated (-). Antibody: Affinity purified rabbit Histone H2AX [p Ser139] antibody NB100-384 used for WB at 0.1 ug/ml. Detection: Chemiluminescence with an exposure time of 3 seconds. Band appears at an observed molecular weight of ~18 kDa.

FFPE section of mouse CT26 colon carcinoma. Antibody: Affinity purified rabbi Histone H2AX [p Ser139] antibody used at a dilution of 1:1,000 (1 ug/ml). Detection: DAB.

Analysis of Histone H2AX [p Ser139] in EPE Treated Jurkat Cells. Cells were treated for 3 hrs in 5ug/ml etoposide, fixed in 1.5% PFA, and permeabilized in 90% Methanol. 1 million cells were stained with 0.5 ug anti-KLH or anti-H2AX NB100-384 and secondary FITC-conjugated goat anti-rabbit (in a 150ul reaction). Black- etosposide treated, anti-KLH; Red- untreated, anti-Histone H2AX [p Ser139]; Blue- etoposide treated , anti-Histone H2AX [p Ser139].

FFPE section of human ovarian carcinoma. Antibody: Affinity purified rabbit Histone H2AX [p Ser139] antibody used at a dilution of 1:5,000 (0.2 ug/ml). Detection: DAB.

Electropherogram image(s) of corresponding Simple Western lane view. Histone H2AX [p Ser139] antibody was used at 5 ug/ml dilution on Jurkat lysate(s).

Immunohistochemistry: Rabbit Polyclonal Histone H2AX [p Ser139] Antibody - Histone H2AX Antibody on mouse cancer tissue. H2AX (Gray) and H2BGFP(Green). Primary antibody dilution: 1:1000 in a 10um slice.

mTORC1/2 activity prevents Cisplatin-induced cell death in MCF-10A cells. (A) Western blot displaying effects on mTOR signaling during a dose escalation of PP242 treatment in MCF-10A cells; (B) Western blot displaying effects of mTOR signaling on a dose escalation of cisplatin treatment in MCF-10A cells; (C) Western blot displaying effects on mTOR signaling and cell death during non-treated, Cisplatin, PP242, and Cisplatin + PP242-treated MCF-10A cells.

ERM expression and localization in SKBR3 breast cancer cells(A) Western blot analysis of ERM levels in SKBR3, HeLa and PC3 cells. SKBR3 breast cancer cells do not express moesin. (B) Colocalization of ezrin/radixin and ErbB2 in SKBR3 cells. 3D-SIM of fixed cells, stained for endogenous ERM and ErbB2, shows a high degree of colocalization between the ezrin/radixin and ErbB2 at the plasma membrane (left panel: max. projection; middle: single plane section; right: single channels of insert). Scale bars: 10 μm. (C) Analysis of protein association in SKBR3 cells by proximity ligation assay (PLA). 2 h treatment with 3 μM GA leads to decreased association of ezrin/ErbB2 and radixin/ErbB2. Data is represented as mean +/− SEM (***P 1.0 mm, >2.0 mm) are shown. Data are expressed as the mean ± SD (n = 3, *p < 0.01 by t-test). Representative data from three independent experiments are shown. c, d hCB LT-HSCs were cultured for 10 days with control MTM protein or MTM-POT1. After 10 days of culture, LT-HSCs were re-isolated and number of TIF was examined. c Immunocytochemical staining of TRF1 (green), 53BP1 (red), and TOTO3 (blue). Scale bar, 2 μm (left). Frequencies of TIFs after 10 days of culture (right). Data are expressed as the mean ± SD (n = 100: control, n = 100: MTM-POT1, *p < 0.01 by t-test). Representative data from 2 independent experiments are shown. d Immunocytochemical staining of TRF1 (green), RPA32 (red), and TOTO3 (blue). Scale bar, 2 μm (left). Frequencies of TIFs after 10 days of culture (right). Data are expressed as the mean ± SD (n = 110–120: control, n = 110: MTM-POT1, *p < 0.01 by t-test). Representative data from two independent experiments are shown Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/28986560), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Cell numbers positively stained for NeuN, DCX, PCNA and caspase-3 in the subgranular region of the dentate gyrus in WT and RanBP9−/− (KO) mice.(A), DAPI-stained brain sections to show the highlighted subgranular zone within the dentate gyrus region of the hippocampus used for cell counts shown in B. (B), Representative brain sections stained with anti-NeuN, anti-DCX, anti-PCNA, anti-capsase-3 and counter stained with DAPI. Cell counting revealed significantly decreased numbers of NeuN positive cells in RanBP9−/− (KO) brains (22%) compared to WT controls. However DCX, PCNA and caspase positive cell numbers were not significantly altered. In each group, n = 3, data presented as mean± SEM. **, p<0.01 by Student’s t-test. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/23840553), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

SAN1 functions independently of the FA pathway and does not affect FA pathway activation. a, b CSAs of HeLa WT and SAN1−/− cells treated with scrambled ctrl siRNA or FANCD2 siRNA, in response to Cisplatin and MMC (N = 3). Statistical significance determined by two-way ANOVA. Error bars denote s.e.m. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. c Immunoblot showing siRNA knockdown of FANCD2 in HeLa WT and SAN1−/− cells. d IF staining of FANCD2 foci in HeLa WT cells and SAN1−/− cells treated with 0.045 μM MMC. e Immunoblot of FANCD2 showing mono-ubiquitylation in HeLa WT and SAN1−/− cells treated with vehicle or 0.045 μM MMC. f, g CSAs of HeLa WT and SAN1−/− cells treated with ctrl or SNM1A siRNA and exposed to Cisplatin or MMC. Statistical significance was determined by two-way ANOVA test. h Immunoblot of SNM1A in HeLa WT and SAN1−/− cells treated with ctrl or SNM1A siRNA Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/29968717), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Ca

Species Glossary

Reactivity: Hu, Mu, Rt, Ca

Applications:

, , , , , ,

Applications: , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-384AF350
Alexa Fluor 405	NB100-384AF405
Alexa Fluor 488	NB100-384AF488
Alexa Fluor 532	NB100-384AF532
Alexa Fluor 594	NB100-384AF594
Alexa Fluor 647	NB100-384AF647
Alexa Fluor 700	NB100-384AF700
Alexa Fluor 750	NB100-384AF750
Allophycocyanin	NB100-384APC
Biotin	NB100-384B
DyLight 350	NB100-384UV
DyLight 405	NB100-384V
DyLight 488	NB100-384G
DyLight 550	NB100-384R
DyLight 594	NB100-384DL594
DyLight 650	NB100-384C
DyLight 680	NB100-384FR
DyLight 755	NB100-384IR
FITC	NB100-384F
HRP	NB100-384H
Janelia Fluor 549	NB100-384JF549
Janelia Fluor 646	NB100-384JF646
PE	NB100-384PE
PerCP	NB100-384PCP
mFluor Violet 450 SE	NB100-384MFV450
mFluor Violet 500 SE	NB100-384MFV500
mFluor Violet 610 SE	NB100-384MFV610

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NB100-56519

DNMT1 Antibody (60B1220.1) - BSA Free

(4 Reviews)

(140 Publications)

0.1 mg

NB100-56519

Immunohistochemistry-Paraffin: DNMT1 Antibody (60B1220.1) [NB100-56519] - Tissue section of human hepatocellular carcinoma at 1:50 dilution.

Flow Cytometry: DNMT1 Antibody (60B1220.1) [NB100-56519] - An intracellular stain was performed on HeLa cells with DNMT1 [60B1220.1] Antibody NB100-56519AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.

Flow Cytometry: DNMT1 Antibody (60B1220.1) [NB100-56519] - Analysis of PE conjugate of NB100-56519. An intracellular stain was performed on HeLa cells with DNMT1 antibody (60B1220.1) NB100-56519PE (blue) and a matched isotype control NBP2-27287PE (orange). Cells were fixed with 4% PFA and then permeablized with 0

Simple Western: DNMT1 Antibody (60B1220.1) [NB100-56519] - Simple Western lane view shows a specific band for Dnmt1 in 0.5 mg/mL of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Immunoprecipitation: DNMT1 Antibody (60B1220.1) [NB100-56519] - The PCNAS228I mutation differentially affects PCNA binding to PIP box-containing proteins. GST-PIP pull down of His-S-PCNAWT or PCNAS228I. Figure shows Coomassie stained gel of representative pull down (top) and anti-PCNA western blot of the same samples diluted 1:20 (bottom). Amount of 'input' loaded for Coomassie is equivalent to 1%, 'Glut. beads' (Glutathione sepharose 4B beads) is equivalent to 25%. Molecular weight markers are indicated. Image collected and cropped by CiteAb from the following publication (//linkinghub.elsevier.com/retrieve/pii/S1568786416304013), licensed under a CC-BY license.

The PCNAS228I mutation differentially affects PCNA binding to PIP box-containing proteins. B) GST-PIP pull down of His-S-PCNAWT or PCNAS228I. Figure shows Coomassie stained gel of representative pull down (top) and anti-PCNA western blot of the same samples diluted 1:20 (bottom). Amount of 'input' loaded for Coomassie is equivalent to 1%, 'Glut. beads' (Glutathione sepharose 4B beads) is equivalent to 25%. Molecular weight markers are indicated. Image collected and cropped by CiteAb from the following publication (//linkinghub.elsevier.com/retrieve/pii/S1568786416304013), licensed under a CC-BY licence.

WA does not alter DNMTs gene and protein expression. (B) Effect of WA on DNMT1, DNMT3A and DNMT3B protein expression in MDA-MB-231 and MCF-7 cells, as compared to to beta-Actin protein levels. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28467815), licensed under a CC-BY licence.

Verification of the expression of the proteins identified.(A) Validation of the SILAC results of selected proteins in immunoblots of protein extracts from the bladder cancer cells analyzed. The results validated the expression levels of proteins identified by the proteomic approach, including differentially and non-differentially expressed candidates. Antibodies displaying a single predominant band at the expected molecular weights were accepted: and alpha-tubulin, was used as the loading control. GSN, Gelsolin; Cul3, Cullin 3; IPO9. Importin 9; EGFR, Epidermal Growth Factor Receptor; NUP133, Nucleoporin 133; HSP70, Heat Shock Protein 70kDa; MCM6, Minichromosome Maintenance Complex Component 6; RCC1, Regulator of Chromosome Condensation 1; BCAS2, Breast Carcinoma Amplified Sequence 2; DNM, Dynamin; NPM, Nucleophosmin; DCTN, Dynactin; CALR, Calreticulin; MAPK, Mitogen-Activated Protein Kinase; DDX21, DEAD (Asp-Glu-Ala-Asp) box polypeptide 21; CDC2: Cell Division Cycle 2; DNMT1, DNA (cytosine-5)-Methyltransferase 1; MSH6, MutS Homolog 6; RAB14, GTPase Rab14; VDAC, Voltage-Dependent Anion Channel; CK18, Cytokeratin 18; CALD, Caldesmon; CD44, CD44 antigen isoform 1 precursor 2; EZR, Ezrin; MSN, Moesin; ANXA2, Annexin A2; CPNE3, Copine 3; FLNA, Filamin A; CAV1, Caveolin 1. Western Blots were scanned and analyzed using alpha-tubulin as normalizing control. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23308193), licensed under a CC-BY licence.

Reactivity:

Hu, Mu, Rt, Po, Bv, Sh

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Bv, Sh

Applications:

, , , , , , , , ,

Applications: , , , , , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Alt Catalog#

IMG-261A

Alt Catalog# IMG-261A

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-56519AF350
Alexa Fluor 405	NB100-56519AF405
Alexa Fluor 488	NB100-56519AF488
Alexa Fluor 532	NB100-56519AF532
Alexa Fluor 594	NB100-56519AF594
Alexa Fluor 647	NB100-56519AF647
Alexa Fluor 700	NB100-56519AF700
Alexa Fluor 750	NB100-56519AF750
Allophycocyanin	NB100-56519APC
Biotin	NB100-56519B
DyLight 350	NB100-56519UV
DyLight 405	NB100-56519V
DyLight 488	NB100-56519G
DyLight 550	NB100-56519R
DyLight 594	NB100-56519DL594
DyLight 650	NB100-56519C
DyLight 680	NB100-56519FR
DyLight 755	NB100-56519IR
FITC	NB100-56519F
HRP	NB100-56519H
Janelia Fluor 549	NB100-56519JF549
Janelia Fluor 646	NB100-56519JF646
PE	NB100-56519PE
mFluor Violet 450 SE	NB100-56519MFV450
mFluor Violet 500 SE	NB100-56519MFV500
mFluor Violet 610 SE	NB100-56519MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-56519SS
Azide and BSA Free	NBP2-80697

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NB100-305

53BP1 Antibody - BSA Free

(6 Reviews)

(139 Publications)

0.1 mg

NB100-305

<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-305] - 53BP1 Antibody [NB100-305] - Exosomes modulate the repair of DNA DSBs in irradiated recipient cells. Representative images of 53BP1 foci in BHY cells 6 hours after 2 Gy and transfer of BHY exosomes isolated 24 hours after irradiation with 0, 3, 6 or 9 Gy (53BP1 foci green, nuclei blue). Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0152213), licensed under a CC-BY license.

Immunohistochemistry: 53BP1 Antibody [NB100-305] - Immunohistochemical staining of placental villi with 53BP1 Antibody (Catalog #NB100-305) at 40X magnification.

Western Blot: 53BP1 Antibody [NB100-305] - Western blot of whole cell lysate (20 ug/lane) from U2Os cells resolved on a 3 to 8 percent trisacetate gel. A band appears at approximately the theoretical molecular weight of 214 kDa.

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody [NB100-305] - HCC1937 cells (human) were treated with PARPi for 96 h. Nuclei are stained in blue (DAPI) and 53BP1 antibody (foci in green). ICC/IF image submitted by a verified customer review.

Flow Cytometry: 53BP1 Antibody [NB100-305] - An intracellular stain was performed on Ntera2 cells with 53BP1 Antibody NB100-305 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody [NB100-305] - Paraformaldehyde (4%) fixed Saos-2 cells (human osteosarcoma cell line) were stained with anti-53BP1 antibody followed by Alexa Fluor 488 secondary antibody. ICC/IF image submitted by a verified customer review.

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody [NB100-305] - Ntera2 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti- NB100-305 at 2 ug/mL overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:1000 dilution for 60 minutes. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse DyLight 550 (Red) at a 1:1000 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Flow Cytometry: 53BP1 Antibody [NB100-305] - An intracellular stain was performed on HeLa cells with 53BP1 Antibody (Catalog #NB100-305AF488) (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.

Inhibition of ACLY exacerbates radiation induced DNA damage in HNSCC cells and impairs homologous recombination. (C,D) Quantification of cells positive for 53BP1 foci (>5 53BP1 foci/nucleus in a single plane of view) in UM-SCC-6 (C) and UT-SCC-5 (D) cells. Shown are mean percentages of three biological replicates. Immunofluorescent representative images of examples of UM-SCC-6 or UT-SCC-5 cells, 24 h post irradiation, with or without 5 uM BMS303141 pre-treatment. Nuclei in blue, 53BP1 foci in red. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31817870), licensed under a CC-BY licence.

Exosomes modulate the repair of DNA DSBs in irradiated recipient cells. (B) Representative images of 53BP1 foci in BHY cells 6 hours after 2 Gy and transfer of BHY exosomes isolated 24 hours after irradiation with 0, 3, 6 or 9 Gy (53BP1 foci green, nuclei blue). Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0152213), licensed under a CC-BY licence.

$RAP1 protects telomeres against DNA damageWestern blotting showing the expression of RAP1 and TRF2 in MRC-5 cells of different population doublings (PD). Senescent cells (PD 72) were left in culture for at least 4 weeks before harvesting for analysis.ChIP analysis of young (PD 30) and senescent (PD 72) MRC-5 cells performed with either anti-RAP1, anti-TRF2, or an IgG antibody. The immunoprecipitated and input products were loaded into a slot blot membrane and hybridized with a telomere probe, and stripped and hybridized to an Alu probe in order to determine unspecific binding. Quantification was performed by normalizing the telomere signal of the immunoprecipitated to that of the input. Data represent mean +/- SD of three biological replicates.Immunofluorescence detection of 53BP1 (red) and a FISH probe staining telomeres (green) in young (PD 26), pre-senescent (PD 66), and senescent (PD 72 + 4 weeks) MRC-5 fibroblasts. The upper graph shows the percentage of telomere co-localizing with 53BP1 (TIFs). Total DNA damage was measured by immunofluorescence detection of 53BP1. The lower graph shows the total number of 53BP1 foci per nucleus. Approximately 40-50 cells were analyzed per replicate and per condition. Bars represent SEM of two biological replicates.Data information: Statistical analyses were performed using Mann-Whitney U-test (*P 0.05, n = 18; group (mean % of cells with more than five 53BP1 ionizing radiation-induced foci +/−SE); unpaired t test]. See text for details. (C) 53BP1 accumulates at infrequent endogenous chromosomal breaks (indicated by asterisks; no ionizing radiation treatment) in presence or absence of TPX2. (D) Cell cycle profiles of control and TPX2 miRNA expressing HeLa cell cultures obtained via flow cytometry (n = 2). Note that the slight ∼5% increase in the G2/M fraction upon TPX2 depletion can not account for the defect in 53BP1 ionizing radiation-induced foci formation exhibited by ∼55% of TPX2 miRNA expressing cells. See text for details. Error bars represent SE in (B) and SDEV in (D). n = # of independent experiments; NS: non-significant, * p<0.05, ** p<0.01, *** p<0.001; IR: ionizing radiation. Image collected and cropped by CiteAb from the following open publication (//dx.plos.org/10.1371/journal.pone.0110994), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

Hypoxia, blood vessel and proliferation staining. IHC staining in the Biopsy 23 and Biopsy 3 respectively for CA-9, EF5, CD31, Ki67 and LYVE1. 15× Magnification. Image collected and cropped by CiteAb from the following open publication (//www.mdpi.com/2072-6694/4/3/821), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt, Bt, Bv, Ca, Nm, Po, Pm, Eq, Fe, Ha, Pm-Or, Pm, Rb, Sh

Species Glossary

Reactivity: Hu, Mu, Rt, Bt, Bv, Ca, Nm, Po, Pm, Eq, Fe, Ha, Pm-Or, Pm, Rb, Sh

Applications:

, , , , , ,

Applications: , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-305AF350
Alexa Fluor 405	NB100-305AF405
Alexa Fluor 488	NB100-305AF488
Alexa Fluor 532	NB100-305AF532
Alexa Fluor 594	NB100-305AF594
Alexa Fluor 647	NB100-305AF647
Alexa Fluor 700	NB100-305AF700
Alexa Fluor 750	NB100-305AF750
Allophycocyanin	NB100-305APC
Biotin	NB100-305B
DyLight 350	NB100-305UV
DyLight 405	NB100-305V
DyLight 488	NB100-305G
DyLight 550	NB100-305R
DyLight 594	NB100-305DL594
DyLight 650	NB100-305C
DyLight 680	NB100-305FR
DyLight 755	NB100-305IR
FITC	NB100-305F
HRP	NB100-305H
Janelia Fluor 549	NB100-305JF549
Janelia Fluor 646	NB100-305JF646
PE	NB100-305PE
PerCP	NB100-305PCP
mFluor Violet 450 SE	NB100-305MFV450
mFluor Violet 500 SE	NB100-305MFV500
mFluor Violet 610 SE	NB100-305MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-305SS

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NBP2-54753

53BP1 Antibody (1285C) - BSA Free

(2 Publications)

0.1 mg

NBP2-54753

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody (1285C) [NBP2-54753] - Neuro2a cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with 53BP1 Antibody at 5 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) (Catalog #NB100-690) was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Flow (Intracellular): 53BP1 Antibody (1285C) [NBP2-54753] - An intracellular stain was performed on HeLa Cells with 53BP1 (1285C) antibody (Catalog #NBP2-54753) (blue) and a matched isotype control MAB1050 (orange). Cells were fixed with 4% paraformaldehyde, following fixation, cells were permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by rabbit IgG APC-conjugated secondary antibody (F0111, R&D Systems).

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody (1285C) [NBP2-54753] - NIH3T3 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-53BP1 Antibody [1285C] NBP2-54753 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:1000 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Flow Cytometry: 53BP1 Antibody (1285C) [NBP2-54753] - An intracellular stain was performed on Ntera2 cells with 53BP1 Antibody [1285C] NBP2-54753 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody (1285C) [NBP2-54753] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with 53BP1 Antibody at 5 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) (Catalog #NB100-690) was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody (1285C) [NBP2-54753] - NIH-3T3 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with 53BP1 Antibody at 5 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) (Catalog #NB100-690) was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody (1285C) [NBP2-54753] - Ntera2 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-53BP1 Antibody [1285C] NBP2-54753 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:1000 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Reactivity:

Hu, Mu, Rt, Po, Bv, Ca, Eq, Fe, Pm

Species Glossary

Reactivity: Hu, Mu, Rt, Po, Bv, Ca, Eq, Fe, Pm

Applications:

, , ,

Applications: , , ,

Host:

Rabbit Monoclonal

Host: Rabbit Monoclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NBP2-54753AF350
Alexa Fluor 405	NBP2-54753AF405
Alexa Fluor 488	NBP2-54753AF488
Alexa Fluor 532	NBP2-54753AF532
Alexa Fluor 594	NBP2-54753AF594
Alexa Fluor 647	NBP2-54753AF647
Alexa Fluor 700	NBP2-54753AF700
Alexa Fluor 750	NBP2-54753AF750
Allophycocyanin	NBP2-54753APC
Biotin	NBP2-54753B
DyLight 350	NBP2-54753UV
DyLight 405	NBP2-54753V
DyLight 488	NBP2-54753G
DyLight 550	NBP2-54753R
DyLight 594	NBP2-54753DL594
DyLight 650	NBP2-54753C
DyLight 680	NBP2-54753FR
DyLight 755	NBP2-54753IR
FITC	NBP2-54753F
HRP	NBP2-54753H
Janelia Fluor 549	NBP2-54753JF549
Janelia Fluor 646	NBP2-54753JF646
PE	NBP2-54753PE
PerCP	NBP2-54753PCP
mFluor Violet 450 SE	NBP2-54753MFV450
mFluor Violet 500 SE	NBP2-54753MFV500
mFluor Violet 610 SE	NBP2-54753MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NBP2-54753SS
Azide and BSA Free	NBP2-80550

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NB100-904

53BP1 Antibody - BSA Free

(4 Reviews)

(120 Publications)

0.1 ml

NB100-904

Immunohistochemistry: 53BP1 Antibody - BSA Free [NB100-904] - Staining of 53BP1 in human renal cancer cells using DAB with hematoxylin counterstain.

Flow Cytometry: 53BP1 Antibody - BSA Free [NB100-904] - An intracellular stain was performed on Ntera2 cells with 53BP1 Antibody NB100-904 (blue) and a matched isotype control NBP2-24891 (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Western Blot: 53BP1 Antibody - BSA Free [NB100-904] - Western blot analysis of untreated U2OS cell lysate blotted with 53BP1 Antibody. The observed molecular weight here is ~220 kDa, and the theoretical molecular weight is 214 kDa.

<b>Biological Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - AR suppression treatment suppresses DNA damage repair. (A) Cells were treated with AR suppression for 24 h followed by 2 Gy of IR treatment. Representative images of phospho-H2A histone family member X ( -H2AX) and p53-binding protein 1 (53BP1) staining 24 h post-IR of LNCaP cells (scale bar 10 um). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31635359/) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - Immunofluorescence of 53BP1 Antibody in HeLa cells. Image from verified customer review.

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - Immunofluorescence of 53BP1 Antibody in C33A cells. Image from verified customer review.

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.5% Triton X-100. The cells were incubated with 53BP1 Antibody conjugated to Alexa Fluor 488 (Catalog #NB100-904AF488) at 10ug/mL for 1 hour at room temperature. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunocytochemistry/Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - Ntera2 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-53BP1 Antibody NB100-904 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:1000 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Flow Cytometry: 53BP1 Antibody - BSA Free [NB100-904] - An intracellular stain was performed on HeLa cells (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to DyLight 550.

Flow Cytometry: 53BP1 Antibody - BSA Free [NB100-904] - An intracellular stain was performed on HeLa cells with 53BP1 Antibody (Catalog #NB100-904AF647) (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Flow Cytometry: 53BP1 Antibody - BSA Free [NB100-904] - An intracellular stain was performed on HeLa cells with 53BP1 Antibody (Catalog #NB100-904C) (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to DyLight 650.

Cellular RER and not enzyme activity against RNA:DNA hybrids correlates with DNA damage and proinflammatory responseAOverexpression of RNase H1 in Rnaseh2b-/- cells restores RNase H activity against RNA:DNA hybrids to 81 +/- 10% of wild-type levels, while overexpression of RNASEH2B restores cellular enzyme activity for cleavage of both RNA:DNA and DRD:DNA substrates (RER). Rnaseh2b-/- MEFs were complemented with Rnaseh1 (+H1), Rnaseh2b (+H2B) or EGFP by retroviral infection. Mean of n = 3 independent experiments +/- SEM.B, CDNA damage is reduced to wild-type levels by complementation with Rnaseh2b but not Rnaseh1, measured by 53BP1 foci formation in detergent-extracted fixed cells. (B) Representative images (scale bar, 10 um). (C) At least 150 cells were counted for each cell line in three independent experiments. Mean +/- SEM, ****P =10 53BP1 foci with the error bars depicting the s.d. (n = 4). e HeLa cells were transfected with siRNA for 8 h; followed by transfection with the GFP plasmids indicated for another 16 h; irradiated with 2 Gy; and after 24 h of recovery, immunostained for gammaH2AX. The graph shows the percentage of 100 GFP-transfected cells in each experiment whose nuclei contained >=10 gammaH2AX foci with the error bars depicting the s.d. (n = 3). *p < 0.05, **p < 0.01, ns (not significant) as determined by Student's t-test. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32303682), licensed under a CC-BY licence.

AR suppression treatment suppresses DNA damage repair. (A) Cells were treated with AR suppression for 24 h followed by 2 Gy of IR treatment. Representative images of phospho-H2A histone family member X (gamma-H2AX) and p53-binding protein 1 (53BP1) staining 24 h post-IR of LNCaP cells (scale bar 10 um). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31635359), licensed under a CC-BY licence.

hWDR76 interaction network and subnuclear localization suggest conservation of Cmr1 function.(c) Human WDR76 localizes into nuclear foci. Twenty-four hours after GFP-WDR76 transfection, U2OS cells were treated with 10 uM MG132 or 1.5 mM MMS for 2 h. Immunofluorescence analyses of 53BP1 were performed with anti-53BP1 antibody. DAPI was used to stain nuclei. Scale bar, 20 um. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25817432), licensed under a CC-BY licence.

NF kappa B upregulates human PINK1 gene expression. (A) NF kappa B increases the endogenous human PINK1 mRNA level. HEK293 cells were transfected with either the NF kappa B expression vector or empty vector pMTF. RT-PCRs were performed using either primers specific to the human PINK1 coding sequence or the human beta -actin coding sequence. (B) NF kappa B increases the endogenous human PINK1 protein levels. HEK293 cells transfected with NF kappa B were harvested 48 h after transfection for protein detection. Cell lysates were run on 10% glycine gel and images were collected by Licor. A significant increase of endogenous PINK1 was observed. (C) The endogenous human PINK1 protein level was dramatically increased by NF kappa B in SH-SY5Y cells. SH-SY5Y cells were transfected with NF kappa B p65 plasmids or the control vector pMTF and then harvested for protein detections. The human PINK1 protein and its proteolysis products ∆1-PINK1 (55kD) and ∆2-PINK1 (45kD) were all increased. beta -actin acted as the internal control. (D) LPS treatment facilitated PINK1 expression in SH-SY5Y cells. Cells were harvested after being treated for 16 h and then subjected to Western blot. The human beta -actin level was served as a control. Quantification was performed by Image J software. Values indicate means ± SEM. n = 3, *p <0.01 by Student’s t-test. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/25108683), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Initial characterization of the OS-9 protein.(A) OS-9 expression in various human cell lines. Equal protein amounts of total cell lysates were used for SDS-PAGE and subsequent Western blotting. For each cell line, two independent samples are shown. Endogenous OS-9 was detected with a polyclonal antibody raised against a peptide corresponding to amino acids 600–667 of isoform 1 of OS-9. (B) Protein stability assay of endogenous OS-9. U2OS cells were treated with the translational inhibitor cycloheximide (100 µM). At indicated time points, whole cell lysates were analysed by immunoblotting. (C) Effect of hypoxia on OS-9 expression. For hypoxia, UT-7 cells were exposed to 1% O2 for 24 h prior to Western blot analysis. To determine any influence of HIF-1 alpha on OS-9 expression under normoxia, cells were incubated with the prolyl hydroxylase inhibitor DMOG (0.5 mM) for 24 h. (D) Protein interaction between OS-9 and PHD2 in vitro. For co-immunoprecipitation, U2OS cells were transiently co-transfected with the plasmids pOS-9-V5 and pPHD2-His, lysed in NP40 buffer, and subjected to immunoisolation with anti-V5 antibody recognizing OS-9 by its V5-tag. OS-9 and its associated proteins were separated by SDS-PAGE and analyzed by Western blot (lane 2). As controls, samples of untransfected (lane 1) cells or cells transfected with a single plasmid (lanes 3–4) were loaded. Representative Western blots are shown for each subfigure. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/21559462), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

PHD3 depletion stabilizes hypoxic p27 expression by increasing p27 half-life. a Cell cycle arrest at G0 and subsequent release shows an increase of p27 expression in siPHD3 exposed cells. b Quantification for p27 expression under PHD3 depletion at indicated time points after cell cycle release in HeLa and 786-O cells. Asterisk indicates significant difference (p < 0,05; n = 3). c Cell cycle arrest at G0 and inhibition of protein synthesis with cycloheximide indicate increased p27 stability in PHD3 depleted HeLa cells. d Quantification of p27 expression using siPHD3 or control at indicated time points. Four independent experiments (± SEM) are shown (p < 0,05; n = 4). e Analysis of p27 stability in 786–0 cells by cycloheximide chase during reoxygenation after 24 h hypoxia demonstrates markedly increased half-life of p27 upon PHD3 depletion Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/26223520), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

$LRRK2 inhibition does not reduce insoluble alpha -synuclein in wildtype hippocampal neurons. a Primary hippocampal neurons from CD1 pups were treated with 30 nM LRRK2 inhibitors PF-475, PF-360 or DMSO as a vehicle control, then transduced with 2.5 μg/mL alpha -synuclein PFFs and allowed to age a further 14 days prior to sequential detergent fractionation. TX-100-insoluble alpha -synuclein and p62 are similar in both LRRK2 inhibitor treated and untreated neurons. b Quantification of soluble proteins show some reduction of LRRK2 protein levels PF-475 treatment and ~ 75% inhibition of LRRK2 activity (as assayed by pS935 LRRK2) by both PF-475 and PF-360. c Insoluble pS129 alpha -synuclein was slightly, but significantly elevated by PF-360 treatment, while alpha -synuclein and p62 were unchanged by one-way ANOVA. (N = 5 biological replicates for each protein). Means + s.e.m.; *P < 0.05, ****p < 0.0001 by one-way ANOVA with Dunnett’s multiple comparison test. All values are normalized first to GAPDH, as a loading control, then to DMSO-treated neurons Image collected and cropped by CiteAb from the following open publication (//actaneurocomms.biomedcentral.com/articles/10.1186/s40478-018-0550-0), licensed under a CC-BY license. Not internally tested by Novus Biologicals.$

TRS functions similarly to eIF4G and acts as an eIF4F analog. a Pull-down assay of co-expressed TRS-Strep with eIF4A- or eIF4G-FLAG in 293 T cells. TRS-Strep was pulled down with Strep-Tactin beads, and co-precipitation of eIF4A or 4 G was determined by immunoblotting with anti-FLAG antibody. EV, empty vector. * indicates a nonspecific band. b Immunoassay of the co-expression of different combinations of plasmids in 293T cells. Myc-TRS was immunoprecipitated with anti-Myc antibody, and co-precipitation of other proteins was determined using tag-specific antibodies. c Immunoassay of co-expressed eIF4A-FLAG with GST-fused full-length TRS or its various domains in 293T cells. eIF4A-FLAG was immunoprecipitated with anti-FLAG antibody, and co-precipitated TRS proteins were determined by immunoblotting with anti-GST antibody. d Pull-down assay of endogenous translation initiation factors with m7GTP-Sepharose beads in 293 T cells transfected with siRNAs against TRS, 4EHP, or a non-targeting control (siCont). Cap-bound proteins were eluted from beads and immunoblotted with the indicated antibodies. Sepharose beads were used as a negative control. e Pull-down assay of endogenous translation initiation factors with m7GTP-Sepharose beads in 293T cells transfected with siRNAs against eIF4G, eIF4E, or siCont, and their suppression effects on cap-binding of other components, were determined as in (d). The data are representative of at least three experiments, each with similar results Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30902983), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

The Neutralizing antibody against nucleolin eliminate HDGF-stimulated AKT/HIF-1 alpha /NF-kappa B/VEGF signaling in SCC4 oral cancer cells. a-d SCC4 cells were treated with recombinant HDGF protein (100 ng/ml) in the presence of anti-NCL or anti-IgG antibody (5 μg/ml) for 24 h before total protein extraction. Cell lysates were subjected to Western blotting with the indicated antibodies. beta -actin was used as an internal control for loading and transfer. Data were mean of three experiments. *, P < 0.05; **, P < 0.01; ns, not statistically significant Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/31711427), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

CIP2A is an inhibitor of PP2A-mediated DAPK dephosphorylationRSC96 cells were transfected with control, CIP2A siRNA (A), DKK-CIP2A (B), or DAPK siRNA (C) for 48 h and then exposed to GAS6 (100 ng/ml) for 30 min. PP2A activity and immunoblotting evaluations of DKK, CIP2A, pDAPK, and DAPK were performed. (D) RSC96 cells were incubated with GAS6 (100 ng/ml) for 30 min and co-immunoprecipitation between CIP2A, pDAPK and PP2A was evaluated. Immunoprecipitated PP2A activity was measured. Data are the mean ± SD, and n = 3 for each time point. * p < 0.05, ** p < 0.01, vs. control. Image collected and cropped by CiteAb from the following open publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.23978), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Other, Pm, Pm-Or, Pm

Species Glossary

Reactivity: Hu, Mu, Other, Pm, Pm-Or, Pm

Applications:

, , , , , ,

Applications: , , , , , ,

Host:

Rabbit Polyclonal

Host: Rabbit Polyclonal

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB100-904AF350
Alexa Fluor 405	NB100-904AF405
Alexa Fluor 488	NB100-904AF488
Alexa Fluor 532	NB100-904AF532
Alexa Fluor 594	NB100-904AF594
Alexa Fluor 647	NB100-904AF647
Alexa Fluor 700	NB100-904AF700
Alexa Fluor 750	NB100-904AF750
Allophycocyanin	NB100-904APC
Allophycocyanin/Cy7	NB100-904APCCY7
Biotin	NB100-904B
DyLight 350	NB100-904UV
DyLight 405	NB100-904V
DyLight 488	NB100-904G
DyLight 550	NB100-904R
DyLight 594	NB100-904DL594
DyLight 650	NB100-904C
DyLight 680	NB100-904FR
DyLight 755	NB100-904IR
FITC	NB100-904F
HRP	NB100-904H
Janelia Fluor 549	NB100-904JF549
Janelia Fluor 646	NB100-904JF646
PE	NB100-904PE
PE/Atto594	NB100-904PEATT594
PE/Cy5.5	NB100-904PECY55
PE/Cy7	NB100-904PECY7
PerCP	NB100-904PCP
mFluor Violet 450 SE	NB100-904MFV450
mFluor Violet 500 SE	NB100-904MFV500
mFluor Violet 610 SE	NB100-904MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB100-904SS

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NB120-13888

DNMT3A Antibody (64B1446) - BSA Free

(3 Reviews)

(121 Publications)

0.1 mg

NB120-13888

Immunohistochemistry-Paraffin: DNMT3A Antibody (64B1446) [NB120-13888] - Detection of DNMT3A on human hepatocellular carcinoma tissue section (NBP2-30221) using 1:100 dilution of DNMT3A antibody (clone 64B1446). The antibody generated a specific cytoplasmic staining in all the cancer cells while some of the cells depicted nuclear staining also.

Immunocytochemistry/Immunofluorescence: DNMT3A Antibody (64B1446) [NB120-13888] - Expression of Dnmt3a in the nuclei of post-mitotic neurons in the olfactory epithelium (OE). DNMT3a (red) is co-expressed with neuron-specific tubulin (green) throughout the development of the olfactory epithelium. Data courtesy of A. Jane Roskams, University of British Columbia.

Flow Cytometry: DNMT3A Antibody (64B1446) [NB120-13888] - Intracellular flow cytometric staining of 1 x 10^6 CHO (A) and HEK-293 (B) cells using Dnmt3a antibody (dark blue). Isotype control shown in orange. An antibody concentration of 1 ug/1x10^6 cells was used.

Flow Cytometry: DNMT3A Antibody (64B1446) [NB120-13888] - Using the PE direct conjugate, an intracellular stain was performed on NTERA-2 cells with DNMT3A (64B1446) NB120-13888PE (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to phycoerythrin.

Immunocytochemistry/Immunofluorescence: DNMT3A Antibody (64B1446) [NB120-13888] - DNMT1 reveals a SCMC-like cortical localization with aberrant localization in Nlrp2tm1a/tm1a derived oocytes. Whole mount immunofluorescence for DNMT3A in unfertilized control oocytes reveals a characteristic metaphase associated localization and no difference is noted in oocytes derived from Nlrp2tm1a/tm1adams. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/srep44667), licensed under a CC-BY license.

Flow Cytometry: DNMT3A Antibody (64B1446) [NB120-13888] - An intracellular stain was performed on Jurkat cells with DNMT3A (64B1446) NB120-13888AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

<b>Biological Strategies Validation. </b>Western Blot: DNMT3A Antibody (64B1446) [NB120-13888] - NTERA-2 cells were treated with Zebularine as indicated for 24 hours. Cell lysates were prepared and separated on a 7.5% gel by SDS-PAGE. Protein was transferred to PVDF membrane and blocked in 5% non-fat milk. The membrane was probed with 2 ug/mL anti-Dnmt3a in 1% milk, and detected with an anti-mouse HRP secondary antibody using chemiluminescence. Note the decrease in Dnmt3a expression upon treatment with 500 uM Zebularine (arrowhead). Additional bands at 90 and 75 kDa can also be detected with this antibody and may represent alternative splice variants. Tubulin is shown as a loading control.

Flow Cytometry: DNMT3A Antibody (64B1446) [NB120-13888] - An intracellular stain was performed on HepG2 cells with DNMT3A (64B1446) NB120-13888APC (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to allophycocyanin (APC).

Flow Cytometry: DNMT3A Antibody (64B1446) [NB120-13888] - An intracellular stain was performed on HepG2 cells with DNMT3A (64B1446) NB120-13888AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Flow Cytometry: DNMT3A Antibody (64B1446) [NB120-13888] - An intracellular stain was performed on HeLa cells with DNMT3A (64B1446) NB120-13888APC (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to allophycocyanin (APC).

<b>Genetic Strategies Validation. </b>Knockdown Validated: DNMT3A Antibody (64B1446) - BSA Free [NB120-13888] - DNMT3A knockdown increases miR‐200b expression via promoter demethylation. Western blot assay of DNMT3A expression in MDA‐MB‐231 and BT549 cells after transfection with DNMT3A siRNA (si‐DNMT3A) or negative control (NC). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30324719/) licensed under a CC-BY license.

DNMT3A knockdown increases miR-200b expression via promoter demethylation. Western blot (C) assay of DNMT3A expression in MDA-MB-231 and BT549 cells after transfection with DNMT3A siRNA (si-DNMT3A) or negative control (NC). Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30324719), licensed under a CC-BY licence.

Protein levels of Dnmt3a and MeCP2 are reduced in inhibitory neurons of conditional knockout mice. (B) Immunofluorescence (IF) images of WT, Dnmt3a cKO or Mecp2 cKO mice probing for Dnmt3a or MeCP2 (red), the Sun1-sfGFP-myc-PA fusion protein that marks the nuclear envelope and is dependent on Cre expression (green), and DAPI to mark genomic DNA (blue). n = 3 mice per genotype (western), n = 3 mice (IF) representative images for two brain regions shown. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32159514), licensed under a CC-BY licence.

Hypoxia and HIF-1 alpha increase GH synthesis.a Effect of hypoxia (1% O2 for 18 h) on GH secretion on nine human acromegalic tumors in primary cell culture. For all cell culture experiments, each GH RIA value was divided to respective cell viability count as determined by WST-1 at OD450nm. Every condition was in quadruplicates and data are means ± SEM. *P < 0.05 to normoxia (NX) (Student’s t test). b Effect of 18 h hypoxia on Gh promoter activity in GH3 cells transfected with the rat Gh-luc plasmid (pA3GHluc). Luc/ beta Gal: luciferase to beta -galactosidase ratio. Data are means ± SEM from three experiments and are presented as percentage of each normoxia (NX). *P < 0.05 to each normoxia (Student’s t test). RLA, relative luciferase activity. c Effect of hypoxia on endogenous rat Gh transcription as determined by real-time RT-PCR. Data are Gh/TfIIb from three independent experiments and presented as fold increase to each normoxia. *P < 0.05 (Student’s t test). d Effect of hypoxia on GH secretion. In b–d transfection with 100 nM HIF-1 alpha siRNA for 48 h abolished the effect of hypoxia. The immunoblot shows as control the HIF-1 alpha siRNA knockdown efficacy in hypoxic cells. Data are means ± SEM from three experiments each done in triplicates. *P < 0.05 to each normoxia (Student’s t test). Effect of HIF-1 alpha overexpression on (e) rat Gh promoter activity, (f) endogenous rat Gh transcription, and (g) secretion. Data are presented as fold increase versus mock control (pCMV empty vector). *P < 0.05 to mock (Student’s t test). h Effect of the HIF-1 alpha mutant HIF-1 alpha R30A on rat Gh promoter activity. Data are means ± SEM from three experiments and are presented as percentage of mock transfected control. Inset shows immunoblot for HIF-1 alpha in the overexpressing cells. i Chromatin immunoprecipitation showing absence of HIF-1 alpha binding to the endogenous rat Gh promoter in GH3 cells under normoxia or hypoxia, in contrast to the positive control Pit-1 (shown in hypoxic cells). Rabbit IgG was used as a control. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32111982), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Reactivity:

Hu, Mu, Rt

Species Glossary

Reactivity: Hu, Mu, Rt

Applications:

, , , , , , , ,

Applications: , , , , , , , ,

Host:

Mouse Monoclonal

Host: Mouse Monoclonal

Alt Catalog#

IMG-268A

Alt Catalog# IMG-268A

Sizes:

Sample Size Available

Sizes: Sample Size Available

Conjugate:

Unconjugated

Available Conjugates:

Validated by:

Validated by:

Conjugate	Catalog #	Availability	Size	Price
Alexa Fluor 350	NB120-13888AF350
Alexa Fluor 405	NB120-13888AF405
Alexa Fluor 488	NB120-13888AF488
Alexa Fluor 532	NB120-13888AF532
Alexa Fluor 594	NB120-13888AF594
Alexa Fluor 647	NB120-13888AF647
Alexa Fluor 700	NB120-13888AF700
Alexa Fluor 750	NB120-13888AF750
Allophycocyanin	NB120-13888APC
Biotin	NB120-13888B
DyLight 350	NB120-13888UV
DyLight 405	NB120-13888V
DyLight 488	NB120-13888G
DyLight 550	NB120-13888R
DyLight 594	NB120-13888DL594
DyLight 650	NB120-13888C
DyLight 680	NB120-13888FR
DyLight 755	NB120-13888IR
FITC	NB120-13888F
HRP	NB120-13888H
Janelia Fluor 549	NB120-13888JF549
Janelia Fluor 646	NB120-13888JF646
PE	NB120-13888PE
mFluor Violet 450 SE	NB120-13888MFV450
mFluor Violet 500 SE	NB120-13888MFV500
mFluor Violet 610 SE	NB120-13888MFV610

Available Formulations

Formulation	Catalog #	Availability	Price
BSA Free	NB120-13888SS
Azide and BSA Free	NBP2-80699

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At	=	A. thaliana	SyHa	=	Golden Syrian Hamster
All	=	All Species	Ha	=	Hamster
An	=	Animal	Hu	=	Human
ArHa	=	Armenian Hamster	I	=	Insect
Av	=	Avian	Ll	=	Llama
Bb	=	Baboon	Ma	=	Mammal
Ba	=	Bacteria	Mk	=	Monkey
Bv	=	Bovine	Mu	=	Mouse
Ca	=	Canine	Multi	=	Multi-species
Ce	=	C. Elegans	NA	=	Non-species specific
Ch	=	Chicken	Or	=	Orangutan
ChHa	=	Chinese Hamster	Pl	=	Plant
Cm	=	Cynomolgus Monkey	Pm	=	Primate
Do	=	Donkey	Po	=	Porcine
Dr	=	Drosophilia	Rb	=	Rabbit
Ec	=	E. Coli	Rt	=	Rat
Eq	=	Equine	RM	=	Rhesus Macaque
Fe	=	Feline	Sh	=	Sheep
Ft	=	Ferret	Vb	=	Vertebrate
Fi	=	Fish	Vi	=	Virus
Fu	=	Fungi	Xp	=	Xenopus
Ge	=	Gerbil	Ye	=	Yeast
GP	=	Guinea Pig	Ze	=	Zebrafish
Gt	=	Goat

Category

Protein/Target

Applications

Reactivity

Citation

Antibody Validation

Reviews

Host

Available Conjugates

Format

Marker

Modification

Clonality

Clone

Gene Symbol

Images Type

Isotype

Kit Type

Lysate Type

Preservative

Protein Or Peptide

Research Areas

Has Sample Size

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Species Glossary

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FANCD2 Antibody - BSA Free

TDP-43/TARDBP Antibody (2E2-D3)

p62/SQSTM1 Antibody - BSA Free

Histone H2AX [p Ser139] Antibody

DNMT1 Antibody (60B1220.1) - BSA Free

53BP1 Antibody - BSA Free

53BP1 Antibody (1285C) - BSA Free

53BP1 Antibody - BSA Free

DNMT3A Antibody (64B1446) - BSA Free