AB76311

Anti-NF-kB p65 antibody [EP2161Y]

Name: Anti-NF-kB p65 antibody [EP2161Y] (ab76311) | Abcam
Brand: Abcam Limited
SKU: AB76311
Price: 205 USD
Availability: InStock
Rating: 4 (1 reviews)

20ul selling size
RabMAb
Recombinant
KO Validated
What is this?

(1 Review)

(30 Publications)

Rabbit Recombinant Monoclonal NF-kB p65 antibody. Suitable for WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples. Cited in 30 publications.

View Alternative Names

NFKB3, RELA, Transcription factor p65, Nuclear factor NF-kappa-B p65 subunit, Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3

5 Images

Immunocytochemistry/ Immunofluorescence - Anti-NF-kB p65 antibody [EP2161Y] (AB76311)

ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-NF-kB p65 antibody [EP2161Y] (AB76311)

Immunocytochemistry/Immunofluorescence analysis HT-29 (human colorectal adenocarcinoma) labelling NF-kB p65 with purified ab76311 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (ab150077). Nuclei counterstained with DAPI (blue).

Control : PBS only

Flow Cytometry (Intracellular) - Anti-NF-kB p65 antibody [EP2161Y] (AB76311)

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-NF-kB p65 antibody [EP2161Y] (AB76311)

Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling NF-kB p65 with purified ab76311 at 1/1000 dilution (1 μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-NF-kB p65 antibody [EP2161Y] (AB76311)

Flow Cytometry analysis of HT-29 (Human colorectal adenocarcinoma epithelial cell) cells labeling NF-kB p65 with purified ab76311 at 1/1000 dilution (1 μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

Supplier Data

Western blot - Anti-NF-kB p65 antibody [EP2161Y] (AB76311)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : NF-κB p65 knockout HAP1 cell lysate (20 μg)
Lane 3 : HeLa cell lysate (20 μg)
Lane 4 : A431 cell lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab76311 observed at 70 kDa. Red - ab8245 loading control, observed at 37 kDa.

ab76311 was shown to specifically react with NF-κB p65 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when NF-κB p65 knockout samples were used. Wild-type and NF-κB p65 knockout samples were subjected to SDS-PAGE. ab76311 (NF-κB p65) and ab8245 (loading control to GAPDH) were both diluted 1/1000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-NF-kB p65 antibody [EP2161Y] (ab76311)

Predicted band size: 60 kDa

false

Unknown

Western blot - Anti-NF-kB p65 antibody [EP2161Y] (AB76311)

All lanes:

Western blot - Anti-NF-kB p65 antibody [EP2161Y] (ab76311) at 1/20000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

Daudi cell lysate at 10 µg

Lane 3:

Jurkat cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 60 kDa

Observed band size: 70 kDa

false

Carrier free

Anti-NF-kB p65 antibody [EP2161Y] - BSA and Azide free
519 Alexa Fluor® 488

Alexa Fluor® 488 Anti-NF-kB p65 antibody [EP2161Y]
665 Alexa Fluor® 647

Alexa Fluor® 647 Anti-NF-kB p65 antibody [EP2161Y]

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EP2161Y

Isotype

IgG

Carrier free

Reacts with

Human

Applications

Flow Cyt (Intra), ICC/IF, WB

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Specificity

ab76311 detects both phosphorylated and non-phosphorylated versions of human NF-kB p65.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000 - 1/5000", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/100 - 1/500", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "1/1000", "FlowCytIntra-species-notes": "<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a> - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody." } } }

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Product details

Species reactivity
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species.
Please contact us for more information.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

NF-kB p65 also known as RelA is a significant component of the NF-kB protein complex. This complex usually involves a molecular weight for p65 of approximately 65 kDa. NF-kB p65 is mechanistically a transcription factor that regulates genes involved in inflammation cell survival and immune response. Expression of p65 is widely seen in various cell types including immune cells and epithelial cells suggesting its role in numerous physiological processes.

Biological function summary

NF-kB p65 acts as part of the larger NF-kB complex usually forming a heterodimer with other family members like p50. This complex translocates to the nucleus upon activation where it binds specific DNA sequences to regulate gene expression. The p65 subunit is essential for transactivating target genes involved in immune and inflammatory responses. Its regulation is important for proper cellular functioning especially in the maintenance of immune homeostasis and inflammatory response.

Pathways

NF-kB p65 participates in critical pathways like the NF-kB signaling and Toll-like receptor pathways. These pathways are fundamental for initiating immune responses and contribute to the regulation of apoptosis and cellular stress responses. In the context of these pathways the IKK complex plays a pivotal role in NF-kB activation leading to the phosphorylation and subsequent degradation of inhibitors that retain NF-kB p65 in the cytoplasm therefore allowing its movement to the nucleus.

NF-kB p65 is implicated in conditions such as cancer and autoimmune diseases. Its dysregulation can lead to persistent activation of inflammatory pathways contributing to tumorigenesis and chronic inflammation. In the context of these diseases proteins such as the p50 subunit and the IKK complex are often involved reflecting the importance of tight regulation of NF-kB signaling in preventing pathological conditions.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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Target data

NF-kappa-B is a pleiotropic transcription factor present in almost all cell types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NF-kappa-B is a homo- or heterodimeric complex formed by the Rel-like domain-containing proteins RELA/p65, RELB, NFKB1/p105, NFKB1/p50, REL and NFKB2/p52. The heterodimeric RELA-NFKB1 complex appears to be most abundant one. The dimers bind at kappa-B sites in the DNA of their target genes and the individual dimers have distinct preferences for different kappa-B sites that they can bind with distinguishable affinity and specificity. Different dimer combinations act as transcriptional activators or repressors, respectively. The NF-kappa-B heterodimeric RELA-NFKB1 and RELA-REL complexes, for instance, function as transcriptional activators. NF-kappa-B is controlled by various mechanisms of post-translational modification and subcellular compartmentalization as well as by interactions with other cofactors or corepressors. NF-kappa-B complexes are held in the cytoplasm in an inactive state complexed with members of the NF-kappa-B inhibitor (I-kappa-B) family. In a conventional activation pathway, I-kappa-B is phosphorylated by I-kappa-B kinases (IKKs) in response to different activators, subsequently degraded thus liberating the active NF-kappa-B complex which translocates to the nucleus. The inhibitory effect of I-kappa-B on NF-kappa-B through retention in the cytoplasm is exerted primarily through the interaction with RELA. RELA shows a weak DNA-binding site which could contribute directly to DNA binding in the NF-kappa-B complex. Besides its activity as a direct transcriptional activator, it is also able to modulate promoters accessibility to transcription factors and thereby indirectly regulate gene expression. Associates with chromatin at the NF-kappa-B promoter region via association with DDX1. Essential for cytokine gene expression in T-cells (PubMed : 15790681). The NF-kappa-B homodimeric RELA-RELA complex appears to be involved in invasin-mediated activation of IL-8 expression. Key transcription factor regulating the IFN response during SARS-CoV-2 infection (PubMed : 33440148).

See full target information RELA

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Publications (30)

Recent publications for all applications. Explore the full list and refine your search

Journal of dental sciences 20:1579-1589 PubMed40654486

2025

A novel NUTM2A-AS1/miR-769-5p axis regulates LPS-evoked damage in human dental pulp cells via the TLR4/MYD88/NF-κB signaling.

Applications

Unspecified application

Species

Unspecified reactive species

Jing Li,Zeru Wang

Allergologia et immunopathologia 53:112-122 PubMed39786883

2025

Acanthoside B attenuates NLRP3-mediated pyroptosis and ulcerative colitis through inhibition of tAGE/RAGE pathway.

Applications

Unspecified application

Species

Unspecified reactive species

Xiaobo He,Chunfang Zhou,Rui Shang,Xiaoyan Wang

Toxics 12: PubMed39195700

2024

Green Tea Polyphenol Nanoparticles Reduce Anxiety Caused by Tobacco Smoking Withdrawal in Rats by Suppressing Neuroinflammation.

Applications

Unspecified application

Species

Unspecified reactive species

Alaa M Hammad,Lujain F Alzaghari,Malek Alfaraj,Vanessa Lux,Suhair Sunoqrot

Open medicine (Warsaw, Poland) 19:20240991 PubMed39091610

2024

SHP-1 mediates cigarette smoke extract-induced epithelial-mesenchymal transformation and inflammation in 16HBE cells.

Applications

Unspecified application

Species

Unspecified reactive species

Quan He,Shuanglan Xu,Xiaomei Ma,Yuanxia Qian,Xuzhi Lu,Weiqi Feng,Zi Chen

Scientific reports 14:17429 PubMed39075107

2024

Platelet-derived exosomes regulate endothelial cell inflammation and M1 macrophage polarization in coronary artery thrombosis via modulating miR-34a-5p expression.

Applications

Unspecified application

Species

Unspecified reactive species

Kangkang Wei,Lintong Yu,Jinming Li,Jie Gao,Li Chen,Min Liu,Xiaohan Zhao,Min Li,Dazhuo Shi,Xiaojuan Ma

Molecules (Basel, Switzerland) 28: PubMed37446793

2023

Chronotoxicity of Acrylamide in Mice Fed a High-Fat Diet: The Involvement of Liver CYP2E1 Upregulation and Gut Leakage.

Applications

Unspecified application

Species

Unspecified reactive species

Luanfeng Wang,Yanhong Liu,Huajing Gao,Shuqi Ge,Xinru Yao,Chang Liu,Xintong Tan

Frontiers in pharmacology 13:1047236 PubMed36699078

2023

Acetylsalicylic acid reduces cigarette smoke withdrawal-induced anxiety in rats modulating the expression of NFĸB, GLT-1, and xCT.

Applications

Unspecified application

Species

Unspecified reactive species

Alaa M Hammad,Lujain F Alzaghari,Malek Alfaraj,Walid Al-Qerem,Wamidh H Talib,Fawaz Alasmari,Haneen Amawi,F Scott Hall

International journal of molecular medicine 50: PubMed35762318

2022

Inhibitory effect of PDE2 on inflammation and apoptosis in cerebral ischemia‑reperfusion injury.

Applications

Unspecified application

Species

Unspecified reactive species

Ding Wan,Xiaoyan Wang,Jin Feng,Peng Wang,Hua Xu

Cell cycle (Georgetown, Tex.) 21:1599-1618 PubMed35404759

2022

Long noncoding RNA BMPR1B-AS1 facilitates endometrial cancer cell proliferation and metastasis by sponging miR-7-2-3p to modulate the DCLK1/Akt/NF-κB pathway.

Applications

Unspecified application

Species

Unspecified reactive species

Tianjiao Lai,Haifeng Qiu,Lulu Si,Yu Zhen,Danxia Chu,Ruixia Guo

Folia neuropathologica 59:335-349 PubMed35114774

2022

Remote ischaemic perconditioning reduces the infarct volume and improves the neurological function of acute ischaemic stroke partially through the miR-153-5p/TLR4/p65/IkBa signalling pathway.

Applications

Unspecified application

Species

Unspecified reactive species

Hao Zha,Wei Miao,Wei Rong,Aimei Wang,Weiqi Jiang,Rui Liu,Lanqi Liu,Ying Wang

View all publications

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Anti-NF-kB p65 antibody [EP2161Y]

Immunocytochemistry/ Immunofluorescence - Anti-NF-kB p65 antibody [EP2161Y] (AB76311)

Flow Cytometry (Intracellular) - Anti-NF-kB p65 antibody [EP2161Y] (AB76311)

Flow Cytometry (Intracellular) - Anti-NF-kB p65 antibody [EP2161Y] (AB76311)

Western blot - Anti-NF-kB p65 antibody [EP2161Y] (AB76311)

Western blot - Anti-NF-kB p65 antibody [EP2161Y] (AB76311)

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Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Specificity

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Reactivity data

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Properties and storage information

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Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage duration

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Aliquoting information

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Publications (30)

Product promise