| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NB100-304SS | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NB100-305SS | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NBP2-54753SS | |||
| Azide and BSA Free | NBP2-80550 | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NB100-904SS | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| BSA Free | NBP2-25028SS | |||
| Azide and BSA Free | NBP2-80549 | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Carrier Free | AF1877-SP | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Carrier Free | AF3405-SP | |||
![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.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-271220231261934.jpg)
![Western Blot: 53BP1 Antibody - BSA Free [NB100-304] - Depletion of TPX2 causes defects in 53BP1 ionizing radiation-induced foci formation.(A) The protein level of 53BP1 is not affected by miRNA-mediated depletion of TPX2. Levels of actin used as loading controls. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0110994), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-310202415334914.jpg)
![Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - Nuclear intensity of key BER proteins.A) Quantification of mean nuclear fluorescence intensity is shown for PARP1 (**** P 25 cells each with Z-stacked (optically sectioned) nuclei.DOI://dx.doi.org/10.7554/eLife.01222.017 Image collected & cropped by CiteAb from the following publication (//elifesciences.org/articles/01222), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-310202415382416.jpg)
![Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - 53BP1 cofractionates with lamins A/C in HDF. (A) HDF were treated with IR (5 Gy, 1 h) & fractionated by sequential treatments of detergent (CSK/T) & DNase I. Extracts were then processed for immunoblotting using the indicated antibodies. WCE, whole cell extract. P, pellet; S, supernatant. (B) HDF were treated with IR (2 Gy; 1 h) & extracted in situ with CSK/T followed by DNase I digestion before being processed for immunofluorescence using lamin A/C & 53BP1 antibodies. Scale bar, 10 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25645366), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-310202415384168.jpg)
![Western Blot: 53BP1 Antibody - BSA Free [NB100-304] - 53BP1 focus formation. (A) Bars indicate the mean ± standard error of 53BP1 focus-forming cells in three independent control & three independent Asciz−/− primary MEF cultures under basal conditions (open bars), in response to 24-h aphidicolin treatment (black bars) & following 30 min recovery after ionizing radiation (grey bars). More detailed quantifications of foci/cell are shown in Supp. Fig. S2. (B) Subcelllular distribution of 53BP1 in cell fractionation assays. M601 is used as a mitochondrial marker, & tubulin as a cytosolic marker. Image collected & cropped by CiteAb from the following publication (//linkinghub.elsevier.com/retrieve/pii/S1568786417301349), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-310202415395939.jpg)
![Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - hsRAD51-II3A expressing cells accumulate 53BP1 foci after treatment with HU. Representative images depicting 53BP1 foci (green) in PCNA (red) positive cells after the indicated treatments. DNA is stained in blue. Scale bar = 10 μm. Dot plot depicts quantitation of 53BP1 foci after indicated treatments. Data is combined data consisting of at least 100 nuclei from two independent experiments. Red line represents the mean. Values are presented as mean ± SD. Statistical significance was determined using the Mann–Whitney U test. Source data are provided as a source data file Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31562309), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-310202415382411.jpg)
![Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - 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), & human normal fibroblast cells (MRC5, BJ). Scale bar: 10 μm. (B) Quantification of (A). Data represent the mean ± SEM, n = 3. More than 100 cells were counted for each experiment. (C) FACS analysis of apoptosis in cells listed in (A). (D) Quantification of (C). Data represent the mean ± SEM, n = 3. (E) Proliferation of p53-positive (VA13, U2OS) & p53-defictive (SAOS2, SKLU-1) ALT cells. (F) Western blot showing activation of JNK (phosphorylation) & accumulation of c-myc in ALT cells. Human normal BJ cells were used as a control Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31115790), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-31020241539594.jpg)
![Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - LNCaP cells in culture were treated for 24 hrs with DMSO or ENZ, then irradiated with 0 or 1 Gy.Cell were fixed & immunostained for 53BP1 0.5, 1.5 & 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. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30933998), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-310202415293335.jpg)
![Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - IMR90 cells depleted for Ajuba undergo a DNA damage response. (A,C,D) Western blots for Ajuba, Cyclin A, Rb, Chk1 phosphorylation, & PARP cleavage on lysates prepared from IMR90 cells 72 h after transfection with siRNA #3. The loading controls with GAPDH for each blot are shown. (B) Staining for p53BP1 in Ajuba-depleted cells (siRNA#3), with siGFP as a control. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23755068), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-310202415371983.jpg)
![Western Blot: 53BP1 Antibody - BSA Free [NB100-304] - 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) Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30559443), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-310202415394519.jpg)
![Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - RNA:DNA hybrids promote 53BP1 foci formation.(a) Immunofluorescence using the anti-flag & anti-53BP1 antibody in cells expressing Flag-tagged hybrid-binding (HB) domain or flag-tagged active RNaseH1. 53BP1 foci were quantified in cells expressing HB-domain or RNaseH1. Column bars represent the mean of n number of cells (described on each column), from three independent experiments. Error bars represent SEM. *P ≤ 0.05; **P 0.05. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25582120), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-310202415525242.jpg)
![Western Blot: 53BP1 Antibody - BSA Free [NB100-304] - Hypoxia causes DNA damage in patient-derived colonosphere cultures in vitro(a) Two independent patient-derived colonospheres were cultured in normoxia (21%) or hypoxia (0.1%) for 24 hours. Immunofluorescence was then used to asses gamma H2AX (green) in the nuclei DAPI (blue). (b) Bar graphs showing the quantification of gamma H2AX observed in (a) (n=3), shown as percentage of the mean gamma H2AX intensity at normoxia. (c) Representative confocal pictures of the comet assay of colonospheres cultured as in (a). (d) Bar graphs showing the quantification of DNA damage observed with the comet assay (Extend tail moment) (n=73). (e) Colonospheres were cultured as in (a) & gamma H2AX was assessed by FACS. The plots show normoxic gamma H2AX levels in red & hypoxic gamma H2AX levels in blue in two independent cell lines. (f) Human colonospheres were exposed to hypoxia for the indicated periods of time. Cell lysates were then analyzed by Western blotting for the indicated markers (cropped). Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.21145), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-31020241683928.jpg)
![Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - TRF1HP1 alpha results in reduced TIFs induced by mutant hTR expression. Cells stably expressing TRF1HP1 alpha (WT or mutant variants of HP1 alpha ) were infected with lentivirus containing WT hTR, or mutant hTR (47A or TSQ1) on day 0, selected for stable expression after 48 h, & analyzed on day 5. a Fluorescence microscopy images of representative cells expressing mutant hTR 47A stained with telomeric (Tel-Cy3) peptide nucleic acid (PNA) probes (magenta in merged image) via fluorescent in situ hybridization (FISH), antibody against DNA-damage repair protein marker 53BP1 (green in merged image), & counterstained with DAPI. Zoom-in images (the last row) correspond to yellow-squared regions of the row above. b % TIF per telomere of each nucleus is quantified; n = 94–159 nuclei combining data of three independent experiments. ****p 0.05. (c) Immunofluorescence staining of gamma H2AX-53BP1 positive foci 7 days following exposure to 1Gy of the indicated radiation type. Scale bar is 10 µm. (d) Immunofluorescence staining for BrdU & 53BP1 7 days following exposure to 1Gy Fe ions. Scale bar is 10 µm. The graph represents the average of scoring in 5 different fields a total of 70 cells. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0108234), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-310202415532010.jpg)
![Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - SPOC1 is recruited to I-SceI-induced DSBs where it colocalizes with 53BP1. (A) In situ immunofluorescence studies with U2OS19 ptight13 GFP-LacR cells with a stably integrated I-SceI cleavage site flanked by lac operator repeats reveals localization of the GFP-lac repressor protein (GFP-LacR) at the lac-operator DNA sequences in the nucleus before (−I-SceI) & 16 h after I-SceI-induced (+ I-SceI) DSB. In contrast, SPOC1 (white) & 53BP1 (red) are distributed throughout the nucleus in the absence of I-Scel, & partially colocalized in naturally occurring repair foci. After I-SceI cleavage to generate DSBs, SPOC1 & 53BP1 colocalize at distinct foci, including the cleaved DNA adjacent to DNA-bound GFP-LacR. Proteins were visualized by immunostaining & confocal microscopy. Scale bars = 10 μm. (B) Quantitative analysis of SPOC1 recruitment to the 53BP1 & GFP-LacR positive lacO array before & 16 h after I-SceI induction. (C) Monitoring of the kinetics of SPOC1 & 53BP1 recruitment to DSBs between 0 & 24 h post-I-SceI induction. (D) Quantifying ATM kinase inhibitor-mediated effects on recruitment of SPOC1 & 53BP1 to DSBs as evident 16 h after I-SceI-induced cleavage. (E) SPOC1 & 53BP1 colocalize at a large discrete endogenous repair focus as observed in some non-irradiated U2OS cells by immunostaining. Scale bar = 10µm. Image collected & cropped by CiteAb from the following publication (//academic.oup.com/nar/article-lookup/doi/10.1093/nar/gks868), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-310202415525292.jpg)
![Western Blot: 53BP1 Antibody - BSA Free [NB100-304] - A-type lamin depletion inhibits DNA repair & reduces cellular fitness to DNA damage. (A) Wild-type (WT) or Y259X HDF treated w/ proteasome inhibitor MG-132 (30 μm). Cell extracts prepared & levels of 53BP1 assessed by immunoblotting. beta -tubulin a loading control. WCE, whole cell extract. The relative levels of 53BP1 expressed relative to levels of untreated WT HDF (1.00). In untreated Y259X cells, 53BP1 levels fell to 15% & restored to ∽50% following treatment w/ MG-132. (B) Cell extracts prepared from WT & Y259X HDF & assessed by immunoblotting w/ indicated antibodies. Shown is a representative set of blots from two independent experiments. (C) WT & Y259X HDF biochemically extracted into cytoplasmic & nuclear fractions before immunoblotting for levels of 53BP1. (D) HDF & Y259X cells treated w/ IR (3 Gy), fixed at the times indicated & stained w/ gamma -H2AX antibody. The number of gamma -H2AX foci at each time point quantified & data represents the mean ± SD from 3 independent experiments. (E) U2OS cells transfected w/ siCTRL or siLMNA siRNA & plated onto 60-mm dishes. The cells then exposed to varying doses of IR & fixed 12 days later in a solution containing crystal violet. The surviving fraction at each IR dose calculated based on the plating efficiency of untreated plates. Data represent the mean ± SD from 3 independent experiments. (F) HDF or Y259X cells treated w/ DMSO or KU55933 ATM inhibitor (10 μm) before exposure to indicated doses of IR then fixed, processed & stained w/ gamma -H2AX antibody. Residual gamma -H2AX foci enumerated & the data represent the mean ± SD from 3 independent experiments. (G) HDF or Y259X cells exposed to IR, briefly extracted w/ CSK buffer & processed for IF w/ gamma -H2AX & pKAP-1 antibodies. Scale bar, 10 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25645366), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-31020241612631.jpg)
![Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - TRF2/Terc double null mutation induces DNA damage response resulting in terminal differentiation of metastatic tumorsHistopathology of primary SCC from K14Cre;TRF2+/+;Terc+/+ A. & K14Cre;TRF2f/f;Terc-/- C. mice is shown by H&E staining. Scale bar = 10 μm. Histopathology of metastatic SCC from K14Cre;TRF2+/+;Terc+/+ B. & K14Cre;TRF2f/f;Terc-/- D., E. mice is shown by H&E staining. Scale bar in (D) = 50 μm. Apoptotic cells in primary SCC from K14Cre;TRF2+/+;Terc+/+ F. & K14Cre;TRF2f/f;Terc-/- G. mice are shown by TUNEL assay. Scale bar = 10 μm. Proliferating cells were detected in K14Cre;TRF2+/+;Terc+/+ H. & K14Cre;TRF2f/f;Terc-/- I. primary SCC by PCNA immunohistochemistry & quantitated in J.. Error bars indicate SEM. Asterisk indicates statistically significant difference. Co-localization of 53BP1 (shown by immunofluorescence, AlexaFluor 488) at telomeres (shown by fluorescence in situ hybridization, Cy3) in histopathologic sections from K14Cre;TRF2+/+;Terc+/+ K. & K14Cre;TRF2f/f;Terc-/- L. primary SCC & quantitated in M. Error bars indicate SEM. Nuclei were counterstained with DAPI. Scale bar = 5 μm. N. Average telomere length ratios in CD34+ stem, Lgr6+ stem, & basal cells from K14Cre;TRF2+/+;Terc+/+ & K14Cre;TRF2f/f;Terc-/- primary SCC were determined by qPCR. Error bars represent SEM. O. Phospho-ATM, phospho-Chk2, & p53 expression in K14Cre;TRF2+/+;Terc+/+ & K14Cre;TRF2f/f;Terc-/- primary SCC is shown by western blot. beta -actin expression was used to control for equal loading of each lane. Representative blots are shown. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/29113290), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-310202415532050.jpg)
![Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - Increased Neurogenesis in the Absence of INK4a/ARF Expression Is Independent of Apoptosis & Microglial Activation(A) Quantification of the number of apoptotic cells, as detected by TUNEL, in the DG 6 hr following exposure of WT, ink4a/arf-deficient (INK4a/ARF KO), & p53-deficient (p53 KO) mice to 6 Gy cranial IR. Indicated is the average number of apoptotic cells per DG section (n = 4 mice per group).(B) Quantification of CD68 expression in the DG 24 hr following exposure of mice to IR.(C) Representative confocal images showing CD68+ cells (in red) next to apoptotic TUNEL+ cells (in green) following exposure of WT mice to IR. Note the increase in CD68 signal & side by side localization of CD68+ & TUNEL+ cells following IR.(D–F) Proportion of neurons (NeuN, in green), young neurons (DCX, in green), & progenitor cells (Sox2, in green) showing persistent DNA damage foci (53BP1, in red) in the hippocampus of mice sacrificed 8 weeks post exposure to IR. Nuclei were stained with DAPI.(G) Representative confocal images of brain sections from irradiated WT mice displaying 53BP1 DNA damage foci with the indicated cell markers are shown. Images were acquired by a Leica microsystems TSC SP8 HyVolution confocal microscope & an HC PL APO CS2 40×/1.3 oil objective. The p values (∗p 25 cells each. (E) GamGFP forms IR-induced foci in Ku80-defective MEFs. (F) Zoomed image from E. (G) IR-induced foci containing Gam only, 53BP1 only or both Gam & 53BP1 (>2600 total foci counted in three independent experiments). Error bars, SEM. Scale bars = 5 μm.DOI://dx.doi.org/10.7554/eLife.01222.012GamGFP marks DSBs in mammalian cells.(A) Live analysis of GamGFP localization to laser-induced DNA damage. Hela cells producing GamGFP were laser damaged along the cell track indicated by the red line at 0 min (m) & images were taken at the indicated times as shown. (B) GamGFP co-localizes with gamma H2AX in fixed Hela cells.DOI://dx.doi.org/10.7554/eLife.01222.013Ku inhibits GamGFP recruitment at DSBs independently of non-homologous end joining.Cells lacking either Ku80 or LigIV are defective in non-homologous end joining (NHEJ), yet the presence of Ku still inhibits recruitment of GamGFP to laser-induced DSBs even in NHEJ-defective cells, & thus independently of the cell's ability to complete NHEJ. Whereas it could have been possible that reduced GamGFP recruitment in the presence of Ku was caused by reduced persistence of DSBs due to their repair by NHEJ, our data show instead that Ku inhibits recruitment independently of successful NHEJ & imply that Ku binding to DSEs itself is inhibitory.DOI://dx.doi.org/10.7554/eLife.01222.014 Image collected & cropped by CiteAb from the following publication (//elifesciences.org/articles/01222), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-310202415525217.jpg)
![Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - Loss of functional PRPF31 induce RNA:DNA hybrid dependent genomic instability but not in mice retinal neurons.(a andb) gamma H2AX & 53BP1 foci analysis in PRPF31 siRNA-transfected RPE-1 cells. (c & d) gamma H2AX & 53BP1 foci analysis in vasculo-stromal fraction derived primary cells fromPrpf31+/A216P mice (Prpf31-ki). (e) gamma H2AX & 53BP1 foci analysis in retina fromPrpf31+/A216P mice on postnatal day 20. All column bars represent the mean. For (a-d) “n”, mentioned on respective column, signify number of cells analyzed from two independent experiments. For (e) n=16 for each column & signify number of retinal sections analyzed; acquired from n=4 eyes. Error bars represent Standard error of Mean (SEM). *P≤0.05; **P 0.9999, ****P 0.05). ns = not significant. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.21145), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-31020241612670.jpg)
![Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - TRF1HP1 alpha results in reduced TIFs induced by mutant hTR expression. Cells stably expressing TRF1HP1 alpha (WT or mutant variants of HP1 alpha ) were infected with lentivirus containing WT hTR, or mutant hTR (47A or TSQ1) on day 0, selected for stable expression after 48 h, & analyzed on day 5. a Fluorescence microscopy images of representative cells expressing mutant hTR 47A stained with telomeric (Tel-Cy3) peptide nucleic acid (PNA) probes (magenta in merged image) via fluorescent in situ hybridization (FISH), antibody against DNA-damage repair protein marker 53BP1 (green in merged image), & counterstained with DAPI. Zoom-in images (the last row) correspond to yellow-squared regions of the row above. b % TIF per telomere of each nucleus is quantified; n = 94–159 nuclei combining data of three independent experiments. ****p 0.05. (c) Immunofluorescence staining of gamma H2AX-53BP1 positive foci 7 days following exposure to 1Gy of the indicated radiation type. Scale bar is 10 µm. (d) Immunofluorescence staining for BrdU & 53BP1 7 days following exposure to 1Gy Fe ions. Scale bar is 10 µm. The graph represents the average of scoring in 5 different fields a total of 70 cells. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0108234), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-31020241682145.jpg)
![Western Blot: 53BP1 Antibody - BSA Free [NB100-304] - Lamin A/C-53BP1 interaction is regulated in a DNA damage-dependent manner. (A) Left; HDF were subjected to IR (10 Gy) & allowed to recover for 1 h. Association between A-type lamins & 53BP1 was assessed by immunoprecipitation of endogenous lamin A/C followed by immunoblotting with 53BP1 & lamin A/C antibodies. WCE, whole cell extract, IP: immunoprecipitates. WCE represents 5% input. Right; endogenous lamin A/C was immunoprecipitated & supernatants or pellets were analysed by immunoblotting for interaction with JMJD2A. (B) U2OS/GFP-lamin A cells were pretreated with caffeine (20 mm) for 1 h before exposure to IR (10 Gy, 1 h recovery). Cell extracts were then subjected to immunoprecipitation using GFP-Trap beads, & bound complexes were then analysed by immunoblotting using 53BP1 & GFP antibodies. WCE represents 1% input. (C) As in (C) except cells were pretreated with 10 μm ATMi for 1 h before IR. WCE represents 1% input. (D) U2OS/GFP-lamin A cells were subjected to laser micro-irradiation, fixed 1 h later & immunostained with gamma -H2AX antibody. Scale bar, 10 μm. (E) U2OS cells were transfected with siCTRL or siLMNA & subjected to laser micro-irradiation, fixed 15 min later & then processed for immunofluorescence with gamma -H2AX & 53BP1 antibodies. Scale bar, 10 μm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25645366), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-310202415525286.jpg)
![Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - Persistence of repair foci at suboptimal pHe. (A) Left panel shows immunofluorescence detection of gamma H2AFX & 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 gamma H2AFX foci/cell at 24 h & 48 h following recovery. Only foci larger than 10 pixels were counted. Experiments were performed in triplicate. Values are mean ± SEM. *P 0.05, n = 18; group (mean % of cells w/ more than five 53BP1 ionizing radiation-induced foci +/−SE); unpaired t test]. See text for details. Image collected & cropped by CiteAb from following publication (//dx.plos.org/10.1371/journal.pone.0110994), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-31020241553744.jpg)
![Western Blot: 53BP1 Antibody - BSA Free [NB100-304] - Aphidicolin-induced ATM signaling in immortalized MEFs. (A) Cells (WT, MJI-53; KO, MJI-1) were treated for 24 h using 0 or 3 μM aphidicolin. (B) Cells (lane 1–3, WT, MJI-91; lanes 4–6, KO, MJI-186; lanes 7–9, WT, MJI-105; lanes 1–12, KO, MJI-84) were treated for 24 h using 0, 1 or 3 μM aphidicolin. Actin & 53BP1 serve as loading controls. DYNLL1 is used as a surrogate marker for loss of ASCIZ. (C) Quantification of western blot band intensities. An arbitrary unit of 100 represents the average band intensity for the respective phospho-protein in the 3 μM aphidicolin-treated wildtype samples on each membrane. Graphs indicate the mean ± standard error, n = 3. Additional loading controls for total KAP1, p53 & H2AX are shown in Supplementary Fig. S1A & B. Image collected & cropped by CiteAb from the following publication (//linkinghub.elsevier.com/retrieve/pii/S1568786417301349), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-310202415532056.jpg)
![Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - POT1 prevents DDR in human HSC. a Expression of POT1 in hCB CD34+ cells & LT-HSCs (Lin–CD34+CD38−CD45RA−CD90+CD49f+). b hCB-derived LT-HSCs were cultured with MTM-POT1. After 10 days of culture, cells were isolated & re-cultured in methylcellulose medium (200 cells per dish). The number of CFU-C & HPP-CFC (>1.0 mm, >2.0 mm) are shown. Data are expressed as the mean ± SD (n = 3, *p 10 foci was called positive & counted). DAPI was used for nuclear staining; (B) The relative DNA damage in primary-culture tumor cells at 120 min post-IR was analyzed by comet assay (upper), & gamma H2AX foci formation in primary-culture tumor cells at 24 h post-IR was presented，“+” & “−“ indicated whether VPA was added in the groups (lower left) & calculated (lower right); (C) The clonogenic survival assay was used to detect survival in the U2OS cells treated with the combination of 0.5 or 1.0 mM VPA with 10 μM ABT888 (PARPi). Each data point in the graphs was from three independent experiments (mean ± SD). p-Values were calculated by Student’s t-test (* p 25 cells each. (E) GamGFP forms IR-induced foci in Ku80-defective MEFs. (F) Zoomed image from E. (G) IR-induced foci containing Gam only, 53BP1 only or both Gam & 53BP1 (>2600 total foci counted in three independent experiments). Error bars, SEM. Scale bars = 5 μm.DOI://dx.doi.org/10.7554/eLife.01222.012GamGFP marks DSBs in mammalian cells.(A) Live analysis of GamGFP localization to laser-induced DNA damage. Hela cells producing GamGFP were laser damaged along the cell track indicated by the red line at 0 min (m) & images were taken at the indicated times as shown. (B) GamGFP co-localizes with gamma H2AX in fixed Hela cells.DOI://dx.doi.org/10.7554/eLife.01222.013Ku inhibits GamGFP recruitment at DSBs independently of non-homologous end joining.Cells lacking either Ku80 or LigIV are defective in non-homologous end joining (NHEJ), yet the presence of Ku still inhibits recruitment of GamGFP to laser-induced DSBs even in NHEJ-defective cells, & thus independently of the cell's ability to complete NHEJ. Whereas it could have been possible that reduced GamGFP recruitment in the presence of Ku was caused by reduced persistence of DSBs due to their repair by NHEJ, our data show instead that Ku inhibits recruitment independently of successful NHEJ & imply that Ku binding to DSEs itself is inhibitory.DOI://dx.doi.org/10.7554/eLife.01222.014 Image collected & cropped by CiteAb from the following publication (//elifesciences.org/articles/01222), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-310202415541936.jpg)
![Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - DNA repair response to irradiation-induced DNA-breaks in adult mice retina.(a) Immunofluorescence performed using anti-gamma H2AX & 53BP1 antibodies in mice retina 1hr after exposure to 5 Gyrase of ionizing radiation. gamma H2AX appears in all cell types in response to DNA breaks. 53BP only observed in the ganglion cell layer (GC) & inner strata of inner nuclear layer (INL) (seeFigure 1a). Zoom in show foci formation in cells expressing 53BP1. (b) H2AX phosphorylation in all retinal cell types is euchromatin specific. (c) Kinetics of DNA repair in mice retina. Post-Irradiation, mice were sacrificed at indicated times & retinal sections were analysed for gamma H2AX & 53BP1. Left panel show all nuclear layers of retina;right panel show zoom in images to emphasize foci formation. Two mice were used for each condition, which were processed & stained together. Random image were taken using confocal microscope on each eye. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30345028), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-310202415541979.jpg)
![Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - Detection of 53BP1 in CTCs from MBC patients in the DETECT IV trial. (a) Selection of MBC patients with CTC-positivity (HER2-) & primary TNBC or primary HER2-, HR+ breast cancer for Eribulin monotherapy & blood sample collection. Blood samples were drawn before therapy (baseline visit), four, twelve, & 24 weeks after the start of Eribulin treatment (1st, 2nd, & 3rd treatment visit) as well as one year after treatment initiation or during premature termination due to disease progression (final visit). The numbers of patients recruited at the different visits are indicated by black columns, the numbers of CTC+ & CTC- patients among them by red & grey columns, respectively. Note that not all of the patients described in Table 1 entered the study during the baseline visit but during later visits which explains the differences in patient numbers. (b) Evaluation of 53BP1 signals in CTCs. Following EpCAM-based CTC enrichment & immunofluorescence microscopic imaging (Cellsearch®), CTCs (CK+, CD45-) were enumerated & 53BP1-staining intensity assessed per individual CTC as indicated by representative examples. A 53BP1 score per patient sample was calculated from the percentage of CTCs without & with light, moderate or strong staining. PBMC, peripheral blood mononuclear cell (CK-, CD45+). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32283863), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-310202415533810.jpg)
![Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - Treatment w/ exogenous POT1 protein protects LT-HSCs in culture. a Effect of MTM-POT1a on colony formation of HSCs. LT-HSCs (8 week-old) cultured w/ MTM-POT1a for 2 weeks. After culture, Lin− cells isolated & re-cultured in MethoCult™ GF M3434 medium (200 cells per dish). Data are expressed as the mean ± SD (n = 3, *p < 0.01 by Tukey’s test). Representative data from two independent experiments are shown. b, c LT-HSCs (8 week-old) cultured w/ MTM-POT1a or control MTM protein. b Number of total cells & LT-HSCs on day 4, 7, & 10 of culture. Data are expressed as the mean ± SD (n = 6, *p < 0.01 by t-test). c Percentage of Annexin V+PI+ apoptotic cells in LT-HSCs on day 4, 7, & 10 of culture. Data are expressed as the mean ± SD (n = 6, *p < 0.01 by t-test). d LT-HSCs (8 week-old) transduced w/ control shRNA or shPot1a-1. After 2 days of shRNA transduction, GFP+LT-HSCs re-sorted & cultured w/ MTM-POT1a or control MTM protein. After 1 week of culture, Annexin V assay performed. Percentage of Annexin V+PI+ apoptotic cells in GFP+LT-HSC fraction is shown. (n = 3, **p < 0.05 by Tukey’s test). e Immunocytochemical staining of TRF (green), 53BP1 (red), & TOTO3 (blue) in 8 week-old LT-HSCs cultured for 3 weeks (left). Frequencies of TIFs after 3 weeks of culture (right). Data are expressed as the mean ± SD (n = 80–100, *p < 0.01 by t-test). f Representative FACS profiles of 8 week-old LT-HSCs after 3 weeks of culture w/ control MTM protein or MTM-POT1a (left). Numbers of LSK cells & LT-HSCs after 3 week culture (starting from 2400 cells) are shown in right panels. Data are expressed as the mean ± SD (n = 3, *p < 0.01 by t-test). Representative data from five independent experiments are shown Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28986560), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-31020241682196.jpg)
![Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-304] - Induction of senescence in hepatocytes causes mitochondrial dysfunction & reduces fatty acid oxidation capacity.Hepatocytes isolated from wild-type C57Bl/6 mice acquire a senescent phenotype 6 days after 10 Gy X-ray irradiation: (a) representative images showing SA-beta -Gal in non-irradiated cells (NOIR) & 6 days after irradiation, Merge: light blue=DAPI, dark blue=SA-beta -Gal, scale bar, 20 μm; (b) frequencies of SA-beta -Gal-positive hepatocytes & (c) representative images showing 53BP1 staining in NOIR & 6 days after irradiation. Merge: blue=DAPI, red=53BP1, scale bar 10 μm; (d) average number of 53BP1 foci per cell are significantly increased 6 days after irradiation. (e) Representative images showing Nile red staining in NOIR & 6 days after IR. Merge: blue=DAPI, red=Nile red, scale bar scale bar 20 μm. (f) Average fluorescence intensity of Nile red staining in senescent or non-senescent hepatocytes. (g) Change in OCR in senescent or non-senescent hepatocytes after addition of the fatty acid palmitate as substrate; (h) change in OCR in senescent or non-senescent hepatocytes after inhibition of fatty acid oxidation by etomoxir. Data in b,g,h are mean±s.e.m. & in d,f are mean±s.d. with 3–4 animals per group. Significant differences (t-test) are indicated with *P≤0.05 & **P≤0.001. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28608850), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-304_rabbit-polyclonal-53bp1-antibody-310202415535184.jpg)
![Immunocytochemistry/Immunofluorescence: 53BP1 Antibody [NB100-305] - 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- 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.](http://images.novusbio.com/fullsize/53BP1-Antibody-Immunocytochemistry-Immunofluorescence-NB100-305-img0010.jpg "Immunocytochemistry/Immunofluorescence: 53BP1 Antibody [NB100-305] - 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- 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.")
![<b>Genetic Strategies Validation. </b>Knockdown Validated: 53BP1 Antibody [NB100-305] - Western blot showing an efficient siRNA-mediated knockdown of the indicated individual or combined proteins. Image collected and cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.11023) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/53BP1-Antibody-Knockdown-Validated-NB100-305-img0013.jpg)
![<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.](http://images.novusbio.com/fullsize/53BP1-Antibody-Immunocytochemistry-Immunofluorescence-NB100-305-img0008.jpg)
![Immunohistochemistry: 53BP1 Antibody [NB100-305] - Immunohistochemical staining of placental villi with 53BP1 Antibody (Catalog #NB100-305) at 40X magnification.](http://images.novusbio.com/fullsize/53BP1-Antibody-Immunohistochemistry-NB100-305-img0005.jpg)
![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.](http://images.novusbio.com/fullsize/53BP1-Antibody-Western-Blot-NB100-305-img0004.jpg)
![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.](http://images.novusbio.com/fullsize/53BP1-Antibody-Immunocytochemistry-Immunofluorescence-NB100-305-img0012.jpg)
![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).](http://images.novusbio.com/fullsize/53BP1-Antibody-Flow-Cytometry-NB100-305-img0011.jpg)
![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.](http://images.novusbio.com/fullsize/53BP1-Antibody-Immunocytochemistry-Immunofluorescence-NB100-305-img0007.jpg)
![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.](http://images.novusbio.com/fullsize/53BP1-Antibody-Immunocytochemistry-Immunofluorescence-NB100-305-img0009.jpg)
![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.](http://images.novusbio.com/fullsize/53BP1-Antibody-Flow-Cytometry-NB100-305-img0006.jpg)
![Western Blot: 53BP1 Antibody - BSA Free [NB100-305] - DNA damage & DNA damage responses upon H2O2 challenge. a, c, e, g Confocal images showing the basal region of the organ of Corti cultures treated with either culture medium alone (a, e) or containing 0.5 mM H2O2 (c, g) for 5 h before being maintained in culture medium alone for 3 days. The samples were then immunolabeled for myosin 7A (red, a, c, e, g), gamma H2AX (green, a & c) & 53BP1 (green, e & g). Scale bars: a, c, e & g = 10 μm. b, d, f, h Higher magnification images of representative OHC & IHC nuclei from all conditions tested. Scale bar = 2.5 μm. i Representative Western blot analysis using antibodies against gamma H2AX, 53BP1, DDB2, p-Chk1, p-Chk2, p53, & beta -actin in whole cochlear extracts. j, k Histograms representing the levels of gamma H2AX, 53BP1, DDB2, p-Chk1, p-Chk2, & p53 in control & in 0.4 & 0.5 mM H2O2-exposed groups (n = 6 cochleae per condition). beta -Actin served as a loading control. Data are expressed as mean ± SEM. One-way ANOVA test followed by post hoc Tukey’s test (*P ≤ 0.04, **P ≤ 0.01, ***P ≤ 0.001 vs. H2O2 0 mM). All experiments were performed in triplicate Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30693443), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-305_rabbit-polyclonal-53bp1-antibody-310202415532062.jpg)
![Western Blot: 53BP1 Antibody - BSA Free [NB100-305] - Assessment of satellite cell radiosensitivity & DNA repair pathways w/ age.(A–B) Expression of major proteins involved in homologous recombination & non-homologous end joining detected using WB. Protein nuclear extracts prepared from activated satellite cells 72 hrs post muscle injury. Lamin B used as loading control. (A) Representative images of WBs. (B) Quantification using Image J software. Data represents mean +/− SEM, n = 3 to 5 (n = a pool of muscles from 3 mice), 2-tailed unpaired Student's t-test, no significant difference. (C) Graphical representation of data from Table S1. Gene expression of old relative to young satellite cells presented as fold change w/ a 95 % confidence interval for the 12 genes w/ a statistically significant relative expression, n = 3 mice per group, two-tailed unpaired Student's t-test, *: p value<0.05. Dotted line represents young level. BER: Base Excision Repair, NER: Nucleotide-Excision Repair; MR: Mismatch Repair, DRR: DNA Repair Related. (D–E) Satellite cells freshly isolated from uninjured (D) or 72 hrs after injury (E) muscle of C57BL/6 young & old mice, & of young SCID mice. Cells plated at low density, irradiated at indicated Gray (Gy) doses, & cultured for 10 days. Myogenic colonies formed by irradiated cells quantified & represented relative to ir respective non-irradiated controls. On average, 178, 77, & 228 myogenic colonies scored per mouse for young, old, & SCID uninjured non-irradiated respectively & 67, 34, & 91 myogenic colonies scored per mouse for young, old, & SCID injured non-irradiated respectively. Data represent mean +/− SEM, n = 3 to 4 mice per group, two-tailed unpaired Student's t-test, a, b or c: p value<0.05, (a: young vs old, b: young vs SCID, c: old versus SCID). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23704914), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-305_rabbit-polyclonal-53bp1-antibody-31020241553204.jpg)
![Immunohistochemistry-Paraffin: 53BP1 Antibody (1285C) [NBP2-54753] - 53BP1 was detected in paraffin-embedded sections of human cervical cancer tissue using Rabbit Anti-Human 53BP1 Monoclonal Antibody (clone 1285C) at 1 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei.](http://images.novusbio.com/fullsize/53BP1-Antibody-1285C-Immunohistochemistry-Paraffin-NBP2-54753-img0004.jpg "Immunohistochemistry-Paraffin: 53BP1 Antibody (1285C) [NBP2-54753] - 53BP1 was detected in paraffin-embedded sections of human cervical cancer tissue using Rabbit Anti-Human 53BP1 Monoclonal Antibody (clone 1285C) at 1 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei.")
![Western Blot: 53BP1 Antibody (1285C) [NBP2-54753] - Total protein from human A431, NTERA-2, HeLa and mouse 3T3 cell lines 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 53BP1 Antibody in block solution and detected with an anti-rabbit HRP secondary antibody using chemiluminescence. The observed molecular weight is shown on this gel at ~250 kDa and the theoretical molecular weight of the whole endogenous protein is 214 kDa.](http://images.novusbio.com/fullsize/53BP1-Antibody-1285C-Western-Blot-NBP2-54753-img0001.jpg)
![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.](http://images.novusbio.com/fullsize/53BP1-Antibody-1285C-Immunocytochemistry-Immunofluorescence-NBP2-54753-img0006.jpg)
![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).](http://images.novusbio.com/fullsize/53BP1-Antibody-1285C-Flow-Intracellular-NBP2-54753-img0008.jpg)
![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.](http://images.novusbio.com/fullsize/53BP1-Antibody-1285C-Immunocytochemistry-Immunofluorescence-NBP2-54753-img0011.jpg)
![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).](http://images.novusbio.com/fullsize/53BP1-Antibody-1285C-Flow-Cytometry-NBP2-54753-img0012.jpg)
![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.](http://images.novusbio.com/fullsize/53BP1-Antibody-1285C-Immunocytochemistry-Immunofluorescence-NBP2-54753-img0005.jpg)
![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.](http://images.novusbio.com/fullsize/53BP1-Antibody-1285C-Immunocytochemistry-Immunofluorescence-NBP2-54753-img0007.jpg)
![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.](http://images.novusbio.com/fullsize/53BP1-Antibody-1285C-Immunocytochemistry-Immunofluorescence-NBP2-54753-img0010.jpg)
![Flow (Intracellular): 53BP1 Antibody (1285C) [NBP2-54753] - An intracellular stain was performed on HeLa cells with 53BP1 [1285C] Antibody (Catalog #NBP2-54753AF647) (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. Image using the Alexa Fluor 647 format of this antibody.](http://images.novusbio.com/fullsize/53BP1-Antibody-1285C-Flow-Intracellular-NBP2-54753-img0009.jpg)
![Western Blot: 53BP1 Antibody - BSA Free [NB100-904] - WRAP53beta plays an important role in recruitment of factors involved in HR and NHEJ to DNA breaks in A2780 cells. A2780 cells were transfected with siControl or siWRAP53#2 oligonucleotides for 48 h; exposed to IR (6 Gy), and 1 h later immunostained for WRAP53beta, gamma-H2AX, MDC1, RNF168, 53BP1, RAD51 and beta-actin. We could not assess the protein levels of BRCA1 due to a lack of antibodies that work for western blotting. The observed molecular weight is 275 kDa and the theoretical molecular weight for the whole endogenous protein is 214 kDa. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/cddis2015250) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/53BP1-Antibody---BSA-Free-Western-Blot-NB100-904-img0015.jpg "Western Blot: 53BP1 Antibody - BSA Free [NB100-904] - WRAP53beta plays an important role in recruitment of factors involved in HR and NHEJ to DNA breaks in A2780 cells. A2780 cells were transfected with siControl or siWRAP53#2 oligonucleotides for 48 h; exposed to IR (6 Gy), and 1 h later immunostained for WRAP53beta, gamma-H2AX, MDC1, RNF168, 53BP1, RAD51 and beta-actin. We could not assess the protein levels of BRCA1 due to a lack of antibodies that work for western blotting. The observed molecular weight is 275 kDa and the theoretical molecular weight for the whole endogenous protein is 214 kDa. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/cddis2015250) licensed under a CC-BY license.")
![Immunocytochemistry/Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - 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- 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.](http://images.novusbio.com/fullsize/53BP1-Antibody---BSA-Free-Immunocytochemistry-Immunofluorescence-NB100-904-img0021.jpg)
![Immunohistochemistry: 53BP1 Antibody - BSA Free [NB100-904] - Staining of 53BP1 in human renal cancer cells using DAB with hematoxylin counterstain.](http://images.novusbio.com/fullsize/53BP1-Antibody---BSA-Free-Immunohistochemistry-NB100-904-img0008.jpg)
![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).](http://images.novusbio.com/fullsize/53BP1-Antibody---BSA-Free-Flow-Cytometry-NB100-904-img0022.jpg)
![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.](http://images.novusbio.com/fullsize/53BP1-Antibody---BSA-Free-Western-Blot-NB100-904-img0011.jpg)
![<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.](http://images.novusbio.com/fullsize/53BP1 Antibody - BSA Free-Immunocytochemistry-Immunofluorescence-NB100-904-img0023.jpg)
![Immunocytochemistry/Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - Immunofluorescence of 53BP1 Antibody in HeLa cells. Image from verified customer review.](http://images.novusbio.com/fullsize/53BP1-Antibody---BSA-Free-Immunocytochemistry-Immunofluorescence-NB100-904-img0009.jpg)
![Immunocytochemistry/Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - Immunofluorescence of 53BP1 Antibody in C33A cells. Image from verified customer review.](http://images.novusbio.com/fullsize/53BP1-Antibody---BSA-Free-Immunocytochemistry-Immunofluorescence-NB100-904-img0012.jpg)
![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.](http://images.novusbio.com/fullsize/53BP1-Antibody---BSA-Free-Immunocytochemistry-Immunofluorescence-NB100-904-img0013.jpg)
![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.](http://images.novusbio.com/fullsize/53BP1-Antibody---BSA-Free-Immunocytochemistry-Immunofluorescence-NB100-904-img0019.jpg)
![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.](http://images.novusbio.com/fullsize/53BP1-Antibody---BSA-Free-Flow-Cytometry-NB100-904-img0014.jpg)
![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.](http://images.novusbio.com/fullsize/53BP1-Antibody---BSA-Free-Flow-Cytometry-NB100-904-img0017.jpg)
![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.](http://images.novusbio.com/fullsize/53BP1-Antibody---BSA-Free-Flow-Cytometry-NB100-904-img0018.jpg)
![Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - Chromatin opening activates DDR via a mechanism that requires catalytically functional LOXL2 but can be independent of DNA damage. d gamma -H2AX & 53BP1 staining & foci quantification are shown after immunofluorescence w/ indicated antibodies in non-replicative conditions. Dot graphs indicate number of gamma -H2AX (left graph) & 53BP1 (right graph) foci in control & LOXL2 KD cells. Image collected & cropped by CiteAb from following publication (//pubmed.ncbi.nlm.nih.gov/31462706), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-904_rabbit-polyclonal-53bp1-antibody-31020241535691.jpg)
![Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - Integrin-associated proteins affected cell cycle & DNA double strand repairA. Western blot & B. densitometric analysis of p53-wildtype A172 & U87MG cells 48 h after PINCH1, ILK or ILKAP knockdown. Non-specific siRNA served as the control. C. Percentage of BrdU positivity as a marker for S-phase cells at different time points after siRNA transfection. Representative images are shown. D. Immunofluorescence staining of 53BP1 (green) & gamma H2AX (red) & E. quantitative analysis of foci numbers in irradiated A172 & U87MG cells after PINCH1, ILK or ILKAP depletion. Nuclei were stained with DAPI (blue). Data are mean ± SD (n = 3; t-test; *P 10 IR-induced foci. Means±S.D. are shown, n=3. *P-value200 cells were counted per experiment). Expression levels of BRCA1 & 53BP1 were assessed by Western Blot on whole cell extracts 4 days after doxycycline treatment. A representative picture is shown in (C). The migration positions of molecular weight standards (in kDa) are indicated on the left. (D) Quantification of 3 Western blot experiments. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0123333), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-904_rabbit-polyclonal-53bp1-antibody-310202416284918.jpg)
![Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - hWDR76 interaction network & subnuclear localization suggest conservation of Cmr1 function.(a) Domain organization of Cmr1 & human WDR76. Human WDR76 shares 29% sequence similarity with Cmr1 from S. cerevisiae. Filled boxes indicate WD40 repeats. NLS, putative nuclear localization signal. (b) Human WDR76 interacts with HELLS, SUGT1, XRCC5, XRCC6 & the CCT–TRiC complex. SILAC-labelled HeLa cells were transfected with GFP-WDR76 or empty vector. GFP-WDR76 & its interacting proteins were enriched using GFP-Trap resin. Proteins were resolved by SDS–PAGE & digested in-gel with trypsin. Peptides were analysed on a quadrupole Orbitrap mass spectrometer. The plot shows log(10) SILAC ratios of proteins associated with GFP-WDR76 compared with background. WDR76 is highlighted in red & several other interactors are also indicated. (c) Human WDR76 localizes into nuclear foci. Twenty-four hours after GFP-WDR76 transfection, U2OS cells were treated with 10 μM 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 μm. (d) Human WDR76 does not co-localize with PCNA. Stably expressing RFP-PCNA U2OS cells were transfected with GFP-WDR76. 24 h after transfection, cells were fixed & stained with DAPI. Scale bar, 20 μm. (e) Model for the role of Cmr1 in promoting replication recovery. See Discussion for details. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/25817432), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-904_rabbit-polyclonal-53bp1-antibody-310202416284913.jpg)
![Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - Overexpression of RNF8 restores repair of DNA double-strand breaks in splicing-deficient cells. (a) U2OS cells were treated with DMSO, pladienolide B or isoginkgetin for 16 h, irradiated (6 Gy, 1 h recovery) 1 h prior to termination of this treatment, & then subjected to western blotting for H2A, RNF8 & beta -actin. (b & c) U2OS cells were transfected with either GFP-RNF8 or GFP-Empty for 2 h, followed by addition of pladienolide B, isoginkgetin or DMSO, incubation for an additional 5 h, irradiation with 6 Gy, fixation 1 h later, & immunostaining for (b) conjugated ubiquitin (FK2 antibody) & (c) 53BP1. The numbers in white indicate the percentage of 100 transfected, i.e, green cells whose nuclei contained >10 IR-induced foci. Means±S.D. are shown, n=3. (d) U2OS cells were treated as above, except that fixation was performed 24 h after irradiation & the immunostaining was for gamma H2AX. Again, the white numbers indicate the percentage of 100 transfected, i.e, green cells whose nuclei contained >10 gamma H2AX foci. Means±S.D. are shown, n=3. (e) The efficiency of HR measured in direct repeated-GFP U2OS cells transfected with I-SceI in combination with either Flag-Empty or Flag-RNF8 for 24 h, followed by addition of DMSO or pladienolide B & incubation for another 24 h. Means±S.D. are shown, n=3. *P200 cells were counted per experiment). Expression levels of BRCA1 & 53BP1 were assessed by Western Blot on whole cell extracts 4 days after doxycycline treatment. A representative picture is shown in (C). The migration positions of molecular weight standards (in kDa) are indicated on the left. (D) Quantification of 3 Western blot experiments. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0123333), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-904_rabbit-polyclonal-53bp1-antibody-310202416284924.jpg)
![Immunocytochemistry/ Immunofluorescence: 53BP1 Antibody - BSA Free [NB100-904] - WRAP53 beta plays an important role in recruitment of factors involved in HR & NHEJ to DNA breaks in A2780 cells. (a) A2780 cells were transfected with siControl or siWRAP53#2 oligonucleotides for 48 h; exposed to IR (6 Gy), & 1 h later immunostained for gamma H2AX, MDC1, RNF168, FK2 (recognizes conjugated ubiquitin), BRCA1, 53BP1 & RAD51. (b) Quantification of the results in (a), as the percentage of 200 cells counted in each experiment whose nuclei contained >10 IR-induced foci. The error bars depict the S.E.M.; n=3, **P<0.01 & ***P<0.001, as determined by Student's t-test. (c) A2780 cells were treated as in (a) & then subjected to western blotting for WRAP53 beta , gamma H2AX, MDC1, RNF168, 53BP1, RAD51 & beta -actin. We could not assess the protein levels of BRCA1 due to a lack of antibodies that work for western blotting Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/26426684), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nb100-904_rabbit-polyclonal-53bp1-antibody-31020241628493.jpg)
![Knockout Validated: 53BP1 Antibody Pack [NB100-926] - 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.](http://images.novusbio.com/fullsize/53BP1-Antibody-Pack-Knockout-Validated-NB100-926-img0020.jpg "<b>Genetic Strategies Validation. </b>Knockout Validated: 53BP1 Antibody Pack [NB100-926] - 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.")
![<b>Genetic Strategies Validation. </b>Western Blot: 53BP1 Antibody Pack [NB100-926] - Western blot analysis of human 53BP1, blotted with 53BP1 antibody (Catalog #NB100-305). Sample: Whole cell lysate (20 ug/lane) from U2Os cells resolved on a 3 to 8% trisacetate gel. A band appears at approximately the theoretical molecular weight of 214 kDa.](http://images.novusbio.com/fullsize/53BP1-Antibody-Pack-Western-Blot-NB100-926-img0011.jpg)
![Immunocytochemistry/Immunofluorescence: 53BP1 Antibody Pack [NB100-926] - 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) using NB100-305. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0152213), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/53BP1-Antibody-Pack-Immunocytochemistry-Immunofluorescence-NB100-926-img0017.jpg)
![Immunohistochemistry: 53BP1 Antibody Pack [NB100-926] - Immunohistochemical analysis of human breast tumors stained with 53BP1 antibody (Catalog #NB100-304). Image from verified customer review.](http://images.novusbio.com/fullsize/53BP1-Antibody-Pack-NB100-926-img0021.jpg)
![Flow Cytometry: 53BP1 Antibody Pack [NB100-926] - 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.](http://images.novusbio.com/fullsize/53BP1-Antibody-Pack-Flow-Cytometry-NB100-926-img0023.jpg)
![Western Blot: 53BP1 Antibody Pack [NB100-926] - Detection of Human [deleted mouse] 53BP1 by Western Blot on Whole cell lysate from U2OS or 293T cells resolved on a 3 to 8% tris-acetate gel using 53BP1 Antibody (Catalog #NB100-304).](http://images.novusbio.com/fullsize/53BP1-Antibody-Pack-Western-Blot-NB100-926-img0009.jpg)
![Immunohistochemistry: 53BP1 Antibody Pack [NB100-926] - Immunohistochemical staining of placental villi with 53BP1 Antibody (Catalog #NB100-305) at 40X magnification.](http://images.novusbio.com/fullsize/53BP1-Antibody-Pack-Immunohistochemistry-NB100-926-img0012.jpg)
![Flow Cytometry: 53BP1 Antibody Pack [NB100-926] - Detection of 53BP1. 1 million Jurkat cells were fixed, permeabilized, and stained with 1.5 mcg/ml 53BP1 Antibody (Catalog #NB100-304) in a 150 mcl reaction. Isotype control (black), anti-53bp1 (red).](http://images.novusbio.com/fullsize/53BP1-Antibody-Pack-Flow-Cytometry-NB100-926-img0010.jpg)
![Flow Cytometry: 53BP1 Antibody Pack [NB100-926] - 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.](http://images.novusbio.com/fullsize/53BP1-Antibody-Pack-Flow-Cytometry-NB100-926-img0022.jpg)
![Immunocytochemistry/Immunofluorescence: 53BP1 Antibody (6B3E10) [NBP2-25028] - Staining of human U-2 OS cells using anti-53BP1 antibody. Bottom images: human U-2 OS cells were treated with EPE to induce DNA-damage and showed increase in nuclear spots after the induction of DNA damage with EPE. Image from verified customer review.](http://images.novusbio.com/fullsize/53BP1-Antibody-6B3E10-Immunocytochemistry-Immunofluorescence-NBP2-25028-img0006.jpg "<b>Independent Antibodies Validation and Biological Strategies Validation.</b>Immunocytochemistry/Immunofluorescence: 53BP1 Antibody (6B3E10) [NBP2-25028] - Staining of human U-2 OS cells using anti-53BP1 antibody. Bottom images: human U-2 OS cells were treated with EPE to induce DNA-damage and showed increase in nuclear spots after the induction of DNA damage with EPE. Image from verified customer review.")
![<b>Independent Antibodies Validation and Biological Strategies Validation.</b>Immunohistochemistry: 53BP1 Antibody (6B3E10) [NBP2-25028] - Analysis of paraffin-embedded colon cancer tissues using 53BP1 Antibody with DAB staining.](http://images.novusbio.com/fullsize/53BP1-Antibody-6B3E10-Immunohistochemistry-NBP2-25028-img0004.jpg)
![Western Blot: 53BP1 Antibody (6B3E10) [NBP2-25028] - Western blot analysis using against HEK293 (1) and 53BP1 (AA: 574-773)-hIgGFc transfected HEK293 (2) cell lysate. The observed molecular weight is ~52 kDa and the theoretical molecular weight of the whole endogenous protein is 214 kDa.](http://images.novusbio.com/fullsize/53BP1-Antibody-6B3E10-Western-Blot-NBP2-25028-img0002.jpg)
![Flow Cytometry: 53BP1 Antibody (6B3E10) [NBP2-25028] - Analysis of HepG2 cells using 53BP1 Antibody (green) and negative control (purple).](http://images.novusbio.com/fullsize/53BP1-Antibody-6B3E10-Flow-Cytometry-NBP2-25028-img0003.jpg)
![Immunohistochemistry: 53BP1 Antibody (6B3E10) [NBP2-25028] - Analysis of paraffin-embedded endometrial cancer tissues using 53BP1 Antibody with DAB staining.](http://images.novusbio.com/fullsize/53BP1-Antibody-6B3E10-Immunohistochemistry-NBP2-25028-img0005.jpg)
![Immunocytochemistry/Immunofluorescence: 53BP1 Antibody (6B3E10) [NBP2-25028] - 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 [6B3E10] NBP2-25028 at 2 ug/ml overnight at 4C and detected with an anti-mouse DyLight 488 (Green) at a 1:1000 dilution for 60 minutes. Beta tubulin NB600-936 was used as a co-stain at a 1:1000 dilution and detected with an anti-rabbit DyLight 550 (Red) at a 1:1000 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.](http://images.novusbio.com/fullsize/53BP1-Antibody-6B3E10-Immunocytochemistry-Immunofluorescence-NBP2-25028-img0007.jpg)
![Flow Cytometry: 53BP1 Antibody (6B3E10) [NBP2-25028] - An intracellular stain was performed on HeLa cells with 53BP1 (6B3E10) Antibody NBP2-25028 (blue) and a matched isotype control MAB002 (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 Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (35503, Thermo Fisher).](http://images.novusbio.com/fullsize/53BP1-Antibody-6B3E10-Flow-Cytometry-NBP2-25028-img0011.jpg)
![Western Blot: 53BP1 Antibody (6B3E10) [NBP2-25028] - Western blot analysis using 53BP1 Antibody against human 53BP1 recombinant protein. The expected molecular weight is 47.6 kDa which is demonstrated in this analysis, and the theoretical molecular weight of the whole endogenous protein is 214 kDa.](http://images.novusbio.com/fullsize/53BP1-Antibody-6B3E10-Western-Blot-NBP2-25028-img0001.jpg)
![Flow Cytometry: 53BP1 Antibody (6B3E10) [NBP2-25028] - An intracellular stain was performed on Ntera2 cells with 53BP1 Antibody [6B3E10] NBP2-25028 (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 1.0 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (35503, Thermo Fisher).](http://images.novusbio.com/fullsize/53BP1-Antibody-6B3E10-Flow-Cytometry-NBP2-25028-img0010.jpg)
![Western Blot: 53BP1 [p Ser25] Antibody [NB100-1803] - Samples: Whole cell lysate (30 ug or 15 ug for WB; 1 mg for IP, 20% of IP loaded) from 293T cells that were mock treated or treated with NCS (200 ng/ml; 1h). Affinity purified rabbit anti-Phospho 53BP1 antibody at 0.04 ug/ml (A) and 0.1 ug/ml (B) for WB. 53BP1 was immunoprecipitated using affinity purified rabbit anti-53BP1 Antibody (Catalog #NB100-304). Detection: Chemiluminescence with exposure times of 3 seconds (A and B). Bands appear at ~350 kDa as the observed molecular weight and the theoretical molecular weight is 214 kDa.](http://images.novusbio.com/fullsize/53BP1-[p-Ser25]-Antibody-Western-Blot-NB100-1803-img0006.jpg "<b>Independent Antibodies Validation and Biological Strategies Validation.</b>Western Blot: 53BP1 [p Ser25] Antibody [NB100-1803] - Samples: Whole cell lysate (30 ug or 15 ug for WB; 1 mg for IP, 20% of IP loaded) from 293T cells that were mock treated or treated with NCS (200 ng/ml; 1h). Affinity purified rabbit anti-Phospho 53BP1 antibody at 0.04 ug/ml (A) and 0.1 ug/ml (B) for WB. 53BP1 was immunoprecipitated using affinity purified rabbit anti-53BP1 Antibody (Catalog #NB100-304). Detection: Chemiluminescence with exposure times of 3 seconds (A and B). Bands appear at ~350 kDa as the observed molecular weight and the theoretical molecular weight is 214 kDa.")
![<b>Independent Antibodies Validation and Biological Strategies Validation.</b>Flow Cytometry: 53BP1 [p Ser25] Antibody [NB100-1803] - Flow cytometric analysis of phospho 53BP1 (S25). Jurkat cells were treated with neocarzinostatin (NCS) for 1.5 hrs. 1 Million cells were fixed, permeabilized, and stained with 1.5 ug/ml antiphospho 53BP1 (S25) (Catalog #NB100-1803) in a 150 ul reaction. Isotype control (black), untreated (red), NCS treated (blue).](http://images.novusbio.com/fullsize/53BP1-[p-Ser25]-Antibody-Flow-Cytometry-NB100-1803-img0002.jpg)
![Immunohistochemistry-Paraffin: 53BP1 [p Ser25] Antibody [NB100-1803] - FFPE sections of human stomach carcinoma (left) and mouse teratoma (right). Antibody: Affinity purified rabbit anti- Phospho 53BP1 (S25) used at a dilution of 1:1,000 (1ug/ml). Detection: DAB.](http://images.novusbio.com/fullsize/53BP1-[p-Ser25]-Antibody-Immunohistochemistry-Paraffin-NB100-1803-img0027.jpg)
![Western Blot: 53BP1 Antibody (HL275) [NBP3-13664] - Non-transfected (-) and transfected (+) HeLa whole cell extracts (30 ug) were separated by 5% SDS-PAGE, and the membrane was blotted with 53BP1 antibody [HL36] (NBP3-13664) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (NBP2-19301) was used to detect the primary antibody.](http://images.novusbio.com/fullsize/53BP1-Antibody-HL275-Western-Blot-NBP3-13664-img0007.jpg "<b>Genetic Strategies Validation. </b>Western Blot: 53BP1 Antibody (HL275) [NBP3-13664] - Non-transfected (-) and transfected (+) HeLa whole cell extracts (30 ug) were separated by 5% SDS-PAGE, and the membrane was blotted with 53BP1 antibody [HL36] (NBP3-13664) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (NBP2-19301) was used to detect the primary antibody.")
![<b>Genetic Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: 53BP1 Antibody (HL275) [NBP3-13664] - 53BP1 antibody [HL275] detects 53BP1 protein at nucleus by immunofluorescent analysis. Sample: HeLa cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: 53BP1 stained by 53BP1 antibody [HL275] (NBP3-13664) diluted at 1:1000. Blue: Fluoroshield with DAPI. Scale bar= 10 um.](http://images.novusbio.com/fullsize/53BP1-Antibody-HL275-Immunocytochemistry-Immunofluorescence-NBP3-13664-img0003.jpg)
![Immunohistochemistry-Paraffin: 53BP1 Antibody (HL275) [NBP3-13664] - 53BP1 antibody [HL275] detects 53BP1 protein at nucleus by immunohistochemical analysis. Sample: Paraffin-embedded human breast carcinoma. 53BP1 stained by 53BP1 antibody [HL275] (NBP3-13664) diluted at 1:200. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min](http://images.novusbio.com/fullsize/53BP1-Antibody-HL275-Immunohistochemistry-Paraffin-NBP3-13664-img0002.jpg)
![Western Blot: 53BP1 Antibody (HL275) [NBP3-13664] - Various whole cell extracts (30 ug) were separated by 5% SDS-PAGE, and the membrane was blotted with 53BP1 antibody [HL36] (NBP3-13664) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (NBP2-19301) was used to detect the primary antibody.](http://images.novusbio.com/fullsize/53BP1-Antibody-HL275-Western-Blot-NBP3-13664-img0001.jpg)
![Western Blot: 53BP1 Antibody (HL275) [NBP3-13664] - Various whole cell extracts (30 ug) were separated by 5% SDS-PAGE, and the membrane was blotted with 53BP1 antibody [HL36] (NBP3-13664) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (NBP2-19301) was used to detect the primary antibody.](http://images.novusbio.com/fullsize/53BP1-Antibody-HL275-Western-Blot-NBP3-13664-img0004.jpg)
![Western Blot: 53BP1 Antibody (HL275) [NBP3-13664] - HeLa whole cell and nuclear extracts (30 ug) were separated by 5% SDS-PAGE, and the membrane was blotted with 53BP1 antibody [HL36] (NBP3-13664) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (NBP2-19301) was used to detect the primary antibody.](http://images.novusbio.com/fullsize/53BP1-Antibody-HL275-Western-Blot-NBP3-13664-img0005.jpg)
![Western Blot: 53BP1 Antibody (HL275) [NBP3-13664] - Various whole cell extracts (30 ug) were separated by 5% SDS-PAGE, and the membrane was blotted with 53BP1 antibody [HL36] (NBP3-13664) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (NBP2-19301) was used to detect the primary antibody.](http://images.novusbio.com/fullsize/53BP1-Antibody-HL275-Western-Blot-NBP3-13664-img0006.jpg)
![Immunohistochemistry-Paraffin: 53BP1 Antibody (HL275) [NBP3-13664] - 53BP1 antibody [HL275] detects 53BP1 protein by immunohistochemical analysis.Sample: Paraffin-embedded rat tissues.53BP1 stained by 53BP1 antibody [HL275] (NBP3-13664) diluted at 1:100.Antigen Retrieval: Citrate buffer, pH 6.0, 15 min](http://images.novusbio.com/fullsize/nbp3-13664_rabbit-53bp1-mab-hl275-91020242011411.jpg)
![Immunohistochemistry-Paraffin: 53BP1 Antibody (HL275) [NBP3-13664] - 53BP1 antibody [HL275] detects 53BP1 protein by immunohistochemical analysis.Sample: Paraffin-embedded mouse tissues.53BP1 stained by 53BP1 antibody [HL275] (NBP3-13664) diluted at 1:100.Antigen Retrieval: Citrate buffer, pH 6.0, 15 min](http://images.novusbio.com/fullsize/nbp3-13664_rabbit-53bp1-mab-hl275-910202420115086.jpg)
![Western Blot: 53BP1 Antibody (HL275) [NBP3-13664] - Various whole cell extracts (30 ug) were separated by 5% SDS-PAGE, and the membrane was blotted with 53BP1 antibody [HL36] (NBP3-13664) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.](http://images.novusbio.com/fullsize/nbp3-13664_rabbit-53bp1-mab-hl275-91020242021972.jpg)
![<b>Orthogonal Strategies Validation. </b>Western Blot: 53BP1 Antibody (HL275) [NBP3-13664] - Various whole cell extracts (30 ug) were separated by 5% SDS-PAGE, and the membrane was blotted with 53BP1 antibody [HL275] diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody. Corresponding RNA expression data for the same cell lines are based on Human Protein Atlas program.](http://images.novusbio.com/fullsize/nbp3-13664_rabbit-53bp1-mab-hl275-1610202419405743.jpg)
![Western Blot: 53BP1 Antibody [NBP2-54677] - Western blot analysis in mouse cell line NIH-3T3 and rat cell line NBT-II. The observed molecular weight as seen on this gel as greater than 250 kDa and the theoretical molecular weight of the whole endogenous protein is 214 kDa.](http://images.novusbio.com/fullsize/53BP1-Antibody-Western-Blot-NBP2-54677-img0007.jpg "Western Blot: 53BP1 Antibody [NBP2-54677] - Western blot analysis in mouse cell line NIH-3T3 and rat cell line NBT-II. The observed molecular weight as seen on this gel as greater than 250 kDa and the theoretical molecular weight of the whole endogenous protein is 214 kDa.")
![Immunocytochemistry/Immunofluorescence: 53BP1 Antibody [NBP2-54677] - Staining of human cell line A-431 with 53BP1 Antibody shows localization to nuclear bodies. Antibody staining is shown in green.](http://images.novusbio.com/fullsize/53BP1-Antibody-Immunocytochemistry-Immunofluorescence-NBP2-54677-img0005.jpg)
![Immunohistochemistry-Paraffin: 53BP1 Antibody [NBP2-54677] - Staining of human cerebellum with 53BP1 Antibody shows strong nuclear positivity.](http://images.novusbio.com/fullsize/53BP1-Antibody-Immunohistochemistry-Paraffin-NBP2-54677-img0001.jpg)
![<b>Genetic Strategies Validation. </b>Western Blot: 53BP1 Antibody [NBP2-54677] - Western blot analysis in U-138MG cells transfected with control siRNA, target specific siRNA probe #1 and #2. Remaining relative intensity is presented. Loading control: Anti-GAPDH. The observed molecular weight as seen on this gel as greater than 250 kDa and the theoretical molecular weight of the whole endogenous protein is 214 kDa.](http://images.novusbio.com/fullsize/53BP1-Antibody-Western-Blot-NBP2-54677-img0006.jpg)
![Western Blot: 53BP1 Antibody [NBP3-14667] - Whole cell lysate (15 ug) from HeLa, HEK293T, MCF-7, Hep-G2, A-549, SW620, SK-MEL-28 (SK), and Jurkat cells prepared using NETN lysis buffer. Rabbit anti-53BP1 recombinant monoclonal antibody [BL-250-1H11] NBP3-14667 lot 1) used at 1:1000. HRP-conjugated goat anti-rabbit IgG (NB7156). Chemiluminescence with an exposure time of 10 second](http://images.novusbio.com/fullsize/53BP1-Antibody-Western-Blot-NBP3-14667-img0001.jpg "Western Blot: 53BP1 Antibody [NBP3-14667] - Whole cell lysate (15 ug) from HeLa, HEK293T, MCF-7, Hep-G2, A-549, SW620, SK-MEL-28 (SK), and Jurkat cells prepared using NETN lysis buffer. Rabbit anti-53BP1 recombinant monoclonal antibody [BL-250-1H11] NBP3-14667 lot 1) used at 1:1000. HRP-conjugated goat anti-rabbit IgG (NB7156). Chemiluminescence with an exposure time of 10 second")
![Immunohistochemistry: 53BP1 Antibody [NBP3-14667] - Sample: FFPE section of mouse renal cell carcinoma.Antibody: Rabbit anti-53BP1 recombinant monoclonalantibody used at 1:250.Secondary: HRP-conjugated goat anti-rabbit IgG. Substrate: DAB.](http://images.novusbio.com/fullsize/53BP1-Antibody-Immunohistochemistry-NBP3-14667-img0004.jpg)
![Immunohistochemistry: 53BP1 Antibody [NBP3-14667] - Sample: FFPE section of colon carcinoma. Antibody: Rabbitanti-53BP1 recombinant monoclonal antibody used at 1:250. Secondary:HRP-conjugated goat anti-rabbit IgG.Substrate: DAB.](http://images.novusbio.com/fullsize/53BP1-Antibody-Immunohistochemistry-NBP3-14667-img0003.jpg)
![Immunoprecipitation: 53BP1 Antibody [NBP3-14667] - Samples: Whole cell lysate (1.0 mgper IP reaction; 20% of IP loaded) from HEK293T cellsprepared using NETN lysis buffer. Antibodies: Rabbitanti-53BP1 recombinant monoclonal antibodyused for IP at 20 ul perreaction. 53BP1 was also immunoprecipitated by rabbitanti-53BP1 antibody for blottingimmunoprecipitated 53BP1, was used at 1:1000.Chemiluminescence with an exposure time of 10 second.](http://images.novusbio.com/fullsize/53BP1-Antibody-Immunoprecipitation-NBP3-14667-img0002.jpg)
