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Rabbit Recombinant Monoclonal NF-kB p65 antibody. Suitable for WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples. Cited in 22 publications.


Images

Western blot - Anti-NF-kB p65 antibody [EP2161Y] (AB76311), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-NF-kB p65 antibody [EP2161Y] (AB76311), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-NF-kB p65 antibody [EP2161Y] (AB76311), expandable thumbnail
  • Western blot - Anti-NF-kB p65 antibody [EP2161Y] (AB76311), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-NF-kB p65 antibody [EP2161Y] (AB76311), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

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

Form
Liquid
Clonality
Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPWBIHC-PICC/IFFlow Cyt (Intra)
Human
Not recommended
Tested
Not recommended
Tested
Tested

Not recommended
Not recommended

Species
Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/1000 - 1/5000
Notes

-

Not recommended
Not recommended

Species
Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/100 - 1/500
Notes

-

Tested
Tested

Species
Human
Dilution info
1/1000
Notes

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Associated Products

Select an associated product type

15 products for Alternative Product

Target data

Function

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. Beside 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).

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

Recommended products

Rabbit Recombinant Monoclonal NF-kB p65 antibody. Suitable for WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples. Cited in 22 publications.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EP2161Y
Purification technique
Affinity purification Protein A
Specificity

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

Concentration
Loading...

Storage

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

Notes

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.

Supplementary info

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

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.

Associated diseases and disorders

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

5 product images

  • Western blot - Anti-NF-kB p65 antibody [EP2161Y] (ab76311), expandable thumbnail

    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 - Anti-GAPDH antibody [6C5] - Loading Control 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 Anti-GAPDH antibody [6C5] - Loading Control 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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

  • Immunocytochemistry/ Immunofluorescence - Anti-NF-kB p65 antibody [EP2161Y] (ab76311), expandable thumbnail

    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 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077). Nuclei counterstained with DAPI (blue).

    Control: PBS only

  • Flow Cytometry (Intracellular) - Anti-NF-kB p65 antibody [EP2161Y] (ab76311), expandable thumbnail

    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, Goat Anti-Rabbit IgG H&L (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).

  • Western blot - Anti-NF-kB p65 antibody [EP2161Y] (ab76311), expandable thumbnail

    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

  • Flow Cytometry (Intracellular) - Anti-NF-kB p65 antibody [EP2161Y] (ab76311), expandable thumbnail

    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, Goat Anti-Rabbit IgG H&L (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).

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Product protocols

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

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