| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Azide and BSA Free | NBP2-27416 | |||
| BSA Free | NB100-56712SS | |||
| Azide and BSA Free | NBP2-27416SS | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Carrier Free | IC5078P | |||
| Carrier Free | IC5078A | |||
| Carrier Free | IC5078G | |||
| Carrier Free | MAB5078-SP | |||
| Carrier Free | IC5078V-100UG | |||
| Carrier Free | IC5078U-100UG | |||
| Carrier Free | IC5078T-100UG | |||
| Carrier Free | IC5078S-100UG | |||
| Carrier Free | IC5078R-100UG | |||
| Carrier Free | IC5078N-100UG | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Carrier Free | AF5078-SP | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Carrier Free | MAB72261-SP | |||
![ELISA: Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric) [NBP2-29661] - Standard curve using the internal RelA standard.](http://images.novusbio.com/fullsize/Human-Mouse-Rat-RelA-NFkB-p65-ELISA-Kit-Colorimetric-ELISA-NBP2-29661-img0004.jpg "ELISA: Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric) [NBP2-29661] - Standard curve using the internal RelA standard.")
![ELISA: Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric) [NBP2-29661] - Nuclear translocation of NF-kB p65 with LPS in Jurkat cells. Jurkat cells, grown to a density of 2X10(6) cells/ml, were treated with LPS (15 ug/ml). The cells were harvested at various indicated time points, and cell lysates were tested in Western analysis using NB100-56712 and also in this ELISA Kit.](http://images.novusbio.com/fullsize/Human-Mouse-Rat-RelA-NFkB-p65-ELISA-Kit-Colorimetric-ELISA-NBP2-29661-img0005.jpg)
![ELISA: Human, Mouse, Rat RelA/NFkB p65 ELISA Kit (Colorimetric) [NBP2-29661] - Immunofluorescence indicates NF-kB p65 subunit was expressed exclusively in AA-G groups and AST-G cytoplasm. High glucose stimulation impact on TSMCs in NF-kB pathway. (A) NF-kB p65 protein subunit expression in TSMCs in each group. (B) NF-kB p65 subunit immunofluorescence. Each number denotes different subunits. Alexa 488 green served as secondary antibodies.Citation: Hara K, Hamada C, Wakabayashi K, Kanda R, Kaneko K, Horikoshi S, et al. (2017) Scavenging of reactive oxygen species by astaxanthin inhibits epithelial-mesenchymal transition in high glucose-stimulated mesothelial cells. PLoS ONE 12(9): e0184332. //doi.org/10.1371/journal.pone.0184332](http://images.novusbio.com/fullsize/Human-Mouse-Rat-RelA-NFkB-p65-ELISA-Kit-Colorimetric-ELISA-NBP2-29661-img0006.jpg)
![Western Blot: RelA/NFkB p65 Antibody [NB100-2176] - Human glomerular endothelial cell lysates, time-response treatment with 10ng/ml TNF-alpha. This image was submitted via customer Review.](http://images.novusbio.com/fullsize/RelA-NFkB-p65-Antibody-Western-Blot-NB100-2176-img0006.jpg "Western Blot: RelA/NFkB p65 Antibody [NB100-2176] - Human glomerular endothelial cell lysates, time-response treatment with 10ng/ml TNF-alpha. This image was submitted via customer Review.")
![Immunohistochemistry-Paraffin: RelA/NFkB p65 Antibody [NB100-2176] - Staining of formalin-fixed, paraffin-embedded human DLBCL showing nuclear expression of NFkB p65 in the tumor cells.](http://images.novusbio.com/fullsize/RelA-NFkB-p65-Antibody-Immunohistochemistry-Paraffin-NB100-2176-img0004.jpg)
![Western Blot: RelA/NFkB p65 Antibody [NB100-2176] - Analysis of NFkB p65 expression in A431 (A) and HeLa (B) whole cell lysates.](http://images.novusbio.com/fullsize/RelA-NFkB-p65-Antibody-Western-Blot-NB100-2176-img0005.jpg)
![Western Blot: RelA/NFkB p65 Antibody (112A1021) [NB100-56712] - Analysis of P65 using p65 antibody at 2 ug/mL in 30 ug of A) Ramos, B) Daudi, C) HeLa and D) mouse NIH 3T3 cell lysate.](http://images.novusbio.com/fullsize/RelA-NFkB-p65-Antibody-112A1021-Western-Blot-NB100-56712-img0013.jpg "Western Blot: RelA/NFkB p65 Antibody (112A1021) [NB100-56712] - Analysis of P65 using p65 antibody at 2 ug/mL in 30 ug of A) Ramos, B) Daudi, C) HeLa and D) mouse NIH 3T3 cell lysate.")
![Simple Western: RelA/NFkB p65 Antibody (112A1021) [NB100-56712] - Lane view shows a specific band for NFkB p65 in 1.0 mg/mL of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.](http://images.novusbio.com/fullsize/RelA-NFkB-p65-Antibody-112A1021-Simple-Western-NB100-56712-img0006.jpg)
![Western Blot: RelA/NFkB p65 Antibody (112A1021) [NB100-56712] - Lysates of HeLa human cervical epithelial carcinoma cell line and Daudi human Burkitt's lymphoma cell line. PVDF membrane was probed with 1 ug/mL mouse anti-RelA/NFkB p65 monoclonal (NB100-56712, Novus Biologicals), followed by 1:2000 dilution of the appropriate HRP-conjugated secondary antibody, donkey anti-mouse IgG (HAF018).](http://images.novusbio.com/fullsize/RelA-NFkB-p65-Antibody-112A1021-Western-Blot-NB100-56712-img0016.jpg)
![Immunohistochemistry-Paraffin: RelA/NFkB p65 Antibody (112A1021) [NB100-56712] - Ovarian cystadenocarcinoma probed with p65 antibody at 5 ug/mL. Human tissue TMA was used for this test. Staining of formalin-fixed tissues is enhanced by boiling tissue sections in 10 mM sodium citrate buffer, pH 6.0 for 10-20 min followed by cooling at RT for 20 min.](http://images.novusbio.com/fullsize/RelA-NFkB-p65-Antibody-112A1021-Immunohistochemistry-Paraffin-NB100-56712-img0009.jpg)
![Flow Cytometry: RelA/NFkB p65 Antibody (112A1021) [NB100-56712] - Intracellular staining of 293 HEK cells using 0.5 ug of p65 antibody. Green histogram represents the isotype control (p65) antibody. This NFkB p65 antibody was used for this test with an anti-mouse IgG PE conjugated secondary antibody.](http://images.novusbio.com/fullsize/RelA-NFkB-p65-Antibody-112A1021-Flow-Cytometry-NB100-56712-img0004.jpg)
![Simple Western: RelA/NFkB p65 Antibody (112A1021) [NB100-56712] - Detection of p65 by Simple Western (JESS) in chemiluminescence in nuclear extract of reconstructed human epidermis lysate (about 600 ug/mL protein concentration).Antibody at 1:20. Simple Western image submitted by a verified customer review.](http://images.novusbio.com/fullsize/RelA-NFkB-p65-Antibody-112A1021-Simple-Western-NB100-56712-img0017.jpg)
![Blocking/Neutralizing: RelA/NFkB p65 [p Ser276] Inhibitor Peptide Set [NBP2-26505] - Effect of various p65-inhibitor peptides on binding of purified recombinant p65 protein to the DNA in vitro. Recombinant p65 protein (100 ng/sample) was incubated for 30 minutes with 50uM of various peptides in 0.025 ml and then assayed for DNA binding activity by EMSA (Please refer to Takada et al, 2004 for further details).](http://images.novusbio.com/fullsize/RelA-NFkB-p65-[p-Ser276]-Inhibitor-Peptide-Set-Blocking-Neutralizing-NBP2-26505-img0008.jpg "Blocking/Neutralizing: RelA/NFkB p65 [p Ser276] Inhibitor Peptide Set [NBP2-26505] - Effect of various p65-inhibitor peptides on binding of purified recombinant p65 protein to the DNA in vitro. Recombinant p65 protein (100 ng/sample) was incubated for 30 minutes with 50uM of various peptides in 0.025 ml and then assayed for DNA binding activity by EMSA (Please refer to Takada et al, 2004 for further details).")
![Blocking/Neutralizing: RelA/NFkB p65 [p Ser276] Inhibitor Peptide Set [NBP2-26505] - PTD-p65-P1 peptide inhibits TNF-induced NF-kB activation: KBM-5 cells were incubated with various concentrations of peptides for 1 hr and treated with 0.1nM TNF for 30 min. Nuclear extracts were prepared and NF-kB activation was analyzed by EMSA (Please refer to Takada et al, 2004 for further details).](http://images.novusbio.com/fullsize/RelA-NFkB-p65-[p-Ser276]-Inhibitor-Peptide-Set-Blocking-Neutralizing-NBP2-26505-img0009.jpg)
![Functional (Inhibition): RelA/NFkB p65 [p Ser276] Inhibitor Peptide Set [NBP2-26505] - TLR5/NF-kB/SEAPorter HEK 293 (NBP2-26277) cells were plated in 96-well plates at 5 x 10^4 cells/well for 16 h. Cells were preincubated with different concentrations (0, 1, 5, 10, 25 and 50 uM) of Inhibitory Peptide (NBP2-26505) and Control Peptide (NBP2-29334) for 1 h. Cells were then stimulated with 1 ng/ml Flagellin (NBP2-25289) for 24 h. Secreted alkaline phosphatase (SEAP) was analyzed using SEAPorter Assay Kit (NBP2-25285). *p < 0.05 versus control peptide at the corresponding concentrations (Mann-Whitney U test).](http://images.novusbio.com/fullsize/RelA-NFkB-p65-p-Ser276-Inhibitor-Peptide-Set-Functional-Inhibition-NBP2-26505-img0010.jpg)
![Western Blot: RelA/NFkB p65 [p Ser536] Antibody [NB100-82088] - HepG2 and HL60 cells untreated or treated with TNF using NFkB-p65(Phospho-Ser536) Antibody.](http://images.novusbio.com/fullsize/RelA-NFkB-p65-[p-Ser536]-Antibody-Western-Blot-NB100-82088-img0013.jpg "Western Blot: RelA/NFkB p65 [p Ser536] Antibody [NB100-82088] - HepG2 and HL60 cells untreated or treated with TNF using NFkB-p65(Phospho-Ser536) Antibody.")
![Immunocytochemistry/Immunofluorescence: RelA/NFkB p65 [p Ser536] Antibody [NB100-82088] - Staining of methanol-fixed Hela cells](http://images.novusbio.com/fullsize/RelA-NFkB-p65-[p-Ser536]-Antibody-Immunocytochemistry-Immunofluorescence-NB100-82088-img0010.jpg)
![Immunohistochemistry-Paraffin: RelA/NFkB p65 [p Ser536] Antibody [NB100-82088] - Analysis of paraffin-embedded human breast carcinoma tissue using NB100-82088 (left) or the same antibody preincubated with blocking peptide (right)](http://images.novusbio.com/fullsize/RelA-NFkB-p65-[p-Ser536]-Antibody-Immunohistochemistry-Paraffin-NB100-82088-img0011.jpg)
![Immunocytochemistry/Immunofluorescence: RelA/NFkB p65 [p Ser536] Antibody [NB100-82088] - Methanol-fixed MEF cells](http://images.novusbio.com/fullsize/RelA-NFkB-p65-[p-Ser536]-Antibody-Immunocytochemistry-Immunofluorescence-NB100-82088-img0009.jpg)
![Immunohistochemistry-Paraffin: RelA/NFkB p65 [p Ser536] Antibody [NB100-82088] - Paraffin-embedded human breast carcinoma tissue.](http://images.novusbio.com/fullsize/RelA-NFkB-p65-[p-Ser536]-Antibody-Immunohistochemistry-Paraffin-NB100-82088-img0007.jpg)
![Immunohistochemistry-Paraffin: RelA/NFkB p65 [p Ser536] Antibody [NB100-82088] - Paraffin-embedded human Lung carcinoma tissue](http://images.novusbio.com/fullsize/RelA-NFkB-p65-[p-Ser536]-Antibody-Immunohistochemistry-Paraffin-NB100-82088-img0008.jpg)
![Western Blot: RelA/NFkB p65 [p Ser536] Antibody [NB100-82088] - Western Blot analysis of various cells using NB100-82088 diluted at 1:2000](http://images.novusbio.com/fullsize/nb100-82088_rabbit-polyclonal-rela-nfkb-p65-p-ser536-antibody-23420241545352.jpg)
![Western Blot: RelA/NFkB p65 [p Ser529, p Ser536] Inhibitor Peptide Set [NBP2-29321] - Inhibits TNF-induced NF-kB activation: KBM-5 cells were incubated with various concentrations of peptides for 1 hr and treated with 0.1nM TNF for 30min. Nuclear extracts were prepared and NF-kB activation was analyzed by EMSA (Please refer toTakada et al, 2004 for further details). PTD-p65-P6 suppresses TNF-induced NF-kB activation by 25% at 100 uM and completely at 150 uM, where as the control peptide, PTD or p65-P6 without PTD did not have any effect.](http://images.novusbio.com/fullsize/RelA-NFkB-p65-[p-Ser529--p-Ser536]-Inhibitor-Peptide-Set-Western-Blot-NBP2-29321-img0015.jpg "Western Blot: RelA/NFkB p65 [p Ser529, p Ser536] Inhibitor Peptide Set [NBP2-29321] - Inhibits TNF-induced NF-kB activation: KBM-5 cells were incubated with various concentrations of peptides for 1 hr and treated with 0.1nM TNF for 30min. Nuclear extracts were prepared and NF-kB activation was analyzed by EMSA (Please refer toTakada et al, 2004 for further details). PTD-p65-P6 suppresses TNF-induced NF-kB activation by 25% at 100 uM and completely at 150 uM, where as the control peptide, PTD or p65-P6 without PTD did not have any effect.")
![Western Blot: RelA/NFkB p65 [p Ser529, p Ser536] Inhibitor Peptide Set [NBP2-29321] - Effect of various p65-inhibitor peptides on binding of purified recombinant p65 protein to the DNA in vitro. Recombinant p65 protein (100 ng/sample) was incubated for 30 minutes with 50uM of various peptides in 0.025 ml and then assayed for DNA binding activity by EMSA (Please refer to Takada et al, 2004 (3) for further details).](http://images.novusbio.com/fullsize/RelA-NFkB-p65-[p-Ser529--p-Ser536]-Inhibitor-Peptide-Set-Western-Blot-NBP2-29321-img0013.jpg)
![Functional (Inhibition): RelA/NFkB p65 [p Ser529, p Ser536] Inhibitor Peptide Set [NBP2-29321] - TLR5/NF-kB/SEAPorter HEK 293 (NBP2-26277) cells were plated in 96-well plates at 5 x 10^4 cells/well for 16 h. Cells were preincubated with different concentrations (0, 1, 5, 10, 25 and 50 uM) of Inhibitory Peptide (NBP2-29321) and Control Peptide (NBP2-29334) for 1 h. Cells were then stimulated with 1 ng/ml Flagellin (NBP2-25289) for 24 h. Secreted alkaline phosphatase (SEAP) was analyzed using SEAPorter Assay Kit (NBP2-25285). *p < 0.05 versus control peptide at the corresponding concentrations (Mann-Whitney U test).](http://images.novusbio.com/fullsize/RelA-NFkB-p65-p-Ser529-p-Ser536-Inhibitor-Peptide-Set-Functional-Inhibition-NBP2-29321-img0012.jpg)
![Western Blot: RelA/NFkB p65 [p Ser276] Antibody [NB100-82086] - Lane 1: HEK293. Lane 2: Lentivirus mediated NFkB overexpression. We image submitted by a verified customer review.](http://images.novusbio.com/fullsize/RelA-NFkB-p65-[p-Ser276]-Antibody-Western-Blot-NB100-82086-img0015.jpg "Western Blot: RelA/NFkB p65 [p Ser276] Antibody [NB100-82086] - Lane 1: HEK293. Lane 2: Lentivirus mediated NFkB overexpression. We image submitted by a verified customer review.")
![Immunocytochemistry/Immunofluorescence: RelA/NFkB p65 [p Ser276] Antibody [NB100-82086] - Immunofluorescence staining of methanol-fixed MEF cells untreated or treated with TNF using NB100-82086](http://images.novusbio.com/fullsize/RelA-NFkB-p65-[p-Ser276]-Antibody-Immunocytochemistry-Immunofluorescence-NB100-82086-img0011.jpg)
![Immunohistochemistry-Paraffin: RelA/NFkB p65 [p Ser276] Antibody [NB100-82086] - Immunohistochemical analysis of paraffin- embedded human lung carcinoma tissue using NB100-82086](http://images.novusbio.com/fullsize/RelA-NFkB-p65-[p-Ser276]-Antibody-Immunohistochemistry-Paraffin-NB100-82086-img0009.jpg)
![Western Blot: RelA/NFkB p65 [p Ser276] Antibody [NB100-82086] - Analysis of extract from HeLa cells using NF-kB p65 (phospho-Ser276) antibody NB100-82086](http://images.novusbio.com/fullsize/RelA-NFkB-p65-[p-Ser276]-Antibody-Western-Blot-NB100-82086-img0012.jpg)
![Immunocytochemistry/Immunofluorescence: RelA/NFkB p65 [p Ser276] Antibody [NB100-82086] - Staining of methanol-fixed Hela cells.](http://images.novusbio.com/fullsize/RelA-NFkB-p65-[p-Ser276]-Antibody-Immunocytochemistry-Immunofluorescence-NB100-82086-img0014.jpg)
![Immunohistochemistry-Paraffin: RelA/NFkB p65 [p Ser276] Antibody [NB100-82086] - Analysis of paraffin- embedded human breast carcinoma tissue using NB100-82086 (left) or the same antibody preincubated with blocking peptide (right)](http://images.novusbio.com/fullsize/RelA-NFkB-p65-[p-Ser276]-Antibody-Immunohistochemistry-Paraffin-NB100-82086-img0013.jpg)
![Immunohistochemistry-Paraffin: RelA/NFkB p65 [p Ser276] Antibody [NB100-82086] - Human breast carcinoma tissue.](http://images.novusbio.com/fullsize/RelA-NFkB-p65-[p-Ser276]-Antibody-Immunohistochemistry-Paraffin-NB100-82086-img0010.jpg)
![Western Blot: RelA/NFkB p65 [p Ser276] Antibody [NB100-82086] - Western Blot analysis of various cells using NB100-82086 diluted at 1:1000](http://images.novusbio.com/fullsize/nb100-82086_rabbit-polyclonal-rela-nfkb-p65-p-ser276-antibody-23420241545358.jpg)
![Western Blot: RelA/NFkB p65 Antibody [NB100-56502] - Analysis of extracts from various cell lines using NFkB p65 antibody at 1:100 (5 ug/mL).](http://images.novusbio.com/fullsize/RelA-NFkB-p65-Antibody-Western-Blot-NB100-56502-img0006.jpg "Western Blot: RelA/NFkB p65 Antibody [NB100-56502] - Analysis of extracts from various cell lines using NFkB p65 antibody at 1:100 (5 ug/mL).")
![Immunocytochemistry/Immunofluorescence: RelA/NFkB p65 Antibody [NB100-56502] - p65 antibody was tested in HeLa cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red).](http://images.novusbio.com/fullsize/RelA-NFkB-p65-Antibody-Immunocytochemistry-Immunofluorescence-NB100-56502-img0004.jpg)
![Western Blot: RelA/NFkB p65 Antibody [NB100-56502] - Analysis of p65 in HeLa cell lysate in the 1) absence and 2) presence of immunizing peptide using this antibody.](http://images.novusbio.com/fullsize/RelA-NFkB-p65-Antibody-Western-Blot-NB100-56502-img0005.jpg)
![Western Blot: RelA/NFkB p65 Antibody (27F9.G4) [NBP1-77815] - RelA/RelA/NFkB p65 p65 antibody (27F9.G4) was used to detect p65 by Western blot. Samples were prepared in RIPA lysis buffer, boiled with NuPage 4x LDS Sample Buffer and run on NuPage 4-12% Bis-Tris Gels. Blot was incubated with primary antibody at a dilution of 1:500 and detected with HRP conjugated anti-mouse antibody at a dilution of 1:10000.](http://images.novusbio.com/fullsize/RelA-NFkB-p65-Antibody-27F9-G4-Western-Blot-NBP1-77815-img0018.jpg "Western Blot: RelA/NFkB p65 Antibody (27F9.G4) [NBP1-77815] - RelA/RelA/NFkB p65 p65 antibody (27F9.G4) was used to detect p65 by Western blot. Samples were prepared in RIPA lysis buffer, boiled with NuPage 4x LDS Sample Buffer and run on NuPage 4-12% Bis-Tris Gels. Blot was incubated with primary antibody at a dilution of 1:500 and detected with HRP conjugated anti-mouse antibody at a dilution of 1:10000.")
![Immunocytochemistry/Immunofluorescence: RelA/NFkB p65 Antibody (27F9.G4) [NBP1-77815] - Tissue: Human Fibroblasts Top: Before activation Bottom: After activation with poly IC Left: anti p65 NLS specific Right: Monoclonal antibody C-Term The two antibodies that are shown target different regions of the p65 protein. The different staining patterns are thought to correspond with different functional regions of the protein.](http://images.novusbio.com/fullsize/RelA-NFkB-p65-Antibody-27F9-G4-Immunocytochemistry-Immunofluorescence-NBP1-77815-img0020.jpg)
![Immunohistochemistry: RelA/NFkB p65 Antibody (27F9.G4) [NBP1-77815] - Antibody has been tested in immunohistochemistry, analyzed by an anatomic pathologist and validated for use in IHC applications against formalin-fixed, paraffin-embedded human tissues. Showed moderate to strong staining within many tissues, including epithelium of the breast, small intestine, kidney, pancreas, prostate, skin, placenta, and uterus, as well as within neurons and lymphoid tissues such as spleen, thymus, and tonsil. The antibody produced an excellent signal with almost no background staining at a concentration of 2.5ug/ml. The image displayed shows specific staining in colon carcinoma as the precipitated red signal, with a hematoxylin purple nuclear counterstain.](http://images.novusbio.com/fullsize/RelA-NFkB-p65-Antibody-27F9-G4-Immunohistochemistry-NBP1-77815-img0019.jpg)
![Western Blot: RelA/NFkB p65 Antibody (27F9.G4) [NBP1-77815] - RelA/NFkB p65 antibody (27F9.G4) was used to detect ~65 kD band (red arrow) in HeLa whole cell lysate. Lysate was run on 4-20% gradient gel transferred under standard conditions and blocked in 1% BSA-TBST for 30 min at RT. Blot was probed with monoclonal anti-NFkB p65 at 1:1000 in 1% BSA-TBST at 4C and detected with HRP conjugated Rb-anti-Mouse antibody at 1:40,000 in for 30 min at RT.](http://images.novusbio.com/fullsize/RelA-NFkB-p65-Antibody-27F9-G4-Western-Blot-NBP1-77815-img0017.jpg)
![Immunocytochemistry/Immunofluorescence: RelA/NFkB p65 Antibody (27F9.G4) [NBP1-77815] - Used at a dilution of 1:5000. Hela cells were grown to sub-confluent on 18 mm2 glass coverslips #1.5. Cells were either unstimulated (A), or stimulated (B) with 50 ng/ml of TNF alpha for 30 min prior fixation. Cells were then fixed in methanol and blocked with 10% normal goat serum (NGS), in PBS, and TritonX 0.2% (Tx) and incubated for 1 hr at RT with primary ab, counterstained with DAPI and washed in PBS/NGS/Tx. Cells were incubated for 1 hr at RT with Atto 425 conjugated anti mouse secondary antibody for STED CW imaging.](http://images.novusbio.com/fullsize/RelA-NFkB-p65-Antibody-27F9-G4-Immunocytochemistry-Immunofluorescence-NBP1-77815-img0012.jpg)
![Immunocytochemistry/Immunofluorescence: RelA/NFkB p65 Antibody (27F9.G4) [NBP1-77815] - Mouse endothelial cells. Unstimulated control cells (left) show cytoplasmic staining, TNF-alpha stimulated cells (right) show nuclear staining. For staining, cells were washed with PBS to remove all traces of culture media and fixed with paraformaldehyde 4 % (45 min). Slides were washed with PBT (phosphate buffer 0.1M + Triton-X-100 0.1 %), 3 times 10 min each, then, blocked with PBT + 5 % normal goat serum, 1 hour. Sample was Incubated overnight in primary antibody (1:600 in blocking buffer). After 3X wash in PBT for 10 min, slides were incubated 1 hour with secondary antibody (1:1000) and mounted in 1:1 PB glycerol.](http://images.novusbio.com/fullsize/RelA-NFkB-p65-Antibody-27F9-G4-Immunocytochemistry-Immunofluorescence-NBP1-77815-img0013.jpg)
![Immunocytochemistry/Immunofluorescence: RelA/NFkB p65 Antibody (27F9.G4) [NBP1-77815] - RelA/NFkB p65 antibody (27F9.G4) was used to detect p65 by immunofluorescence at a dilution of 1:5000. Hela cells were grown to sub-confluent on 18 mm2 glass coverslips #1.5. Cells were either unstimulated (A), or stimulated (B) with 50 ng/ml of TNF alpha for 30 min prior fixation. Cells were then fixed in methanol and blocked with 10% normal goat serum (NGS), in PBS, and TritonX 0.2% (Tx) and incubated for 1 hr at RT with primary ab, counterstained with DAPI and washed in PBS/NGS/Tx. Cells were incubated for 1 hr at RT with Atto 425 conjugated anti mouse secondary antibody for STED CW imaging. Data was collected on a STED-CW TCS-SP5 Confocal system equipped with a DFC 350FX camera allowing sequential acquisition in widefiled, confocal and STED CW imaging on the same system.](http://images.novusbio.com/fullsize/RelA-NFkB-p65-Antibody-27F9-G4-Immunocytochemistry-Immunofluorescence-NBP1-77815-img0015.jpg)
