Anti-NFkB p65 antibody [E379] (ab32536)
- Product nameAnti-NFkB p65 antibody [E379]See all NFkB p65 primary antibodies ...
- DescriptionRabbit monoclonal [E379] to NFkB p65
- Specificityab32536 recognises NF-?B p65.
- Tested applicationsWB, IHC-P, ICC/IF, Flow Cyt, IP more details
- Species reactivityReacts with: Human
Does not react withMouse, Rat
A synthetic peptide corresponding to residues near the C-terminus of human nuclear factor NF-kappa-B p65 subunit
- Positive controlHela cell lysate Human Breast carcinoma HeLa cells
- General notesProduced under U.S. Patent No. 5,675,063.
- Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
- Storage bufferPBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
- Concentration information loading...
- PurityIgG fraction
- Clonality Monoclonal
- Clone numberE379
Our Abpromise guarantee covers the use of ab32536 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||WB: 1/50000 - 1/100000. Detects a band of approximately 65 kDa (predicted molecular weight: 65 kDa).|
|IHC-P||IHC-P: Use at an assay dependent dilution.|
|ICC/IF||ICC/IF: 1/250 - 1/500.|
|Flow Cyt||Flow Cyt: 1/100.|
- FunctionNF-kappa-B is a pleiotropic transcription factor which is present in almost all cell types and is involved in many biological processed 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 and the heterodimeric p65-p50 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. 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. NF-kappa-B heterodimeric p65-p50 and p65-c-Rel complexes are transcriptional activators. The NF-kappa-B p65-p65 complex appears to be involved in invasin-mediated activation of IL-8 expression. The inhibitory effect of I-kappa-B upon NF-kappa-B the cytoplasm is exerted primarily through the interaction with p65. p65 shows a weak DNA-binding site which could contribute directly to DNA binding in the NF-kappa-B complex. Associates with chromatin at the NF-kappa-B promoter region via association with DDX1.
- Sequence similaritiesContains 1 RHD (Rel-like) domain.
- Domainthe 9aaTAD motif is a transactivation domain present in a large number of yeast and animal transcription factors.
modificationsUbiquitinated, leading to its proteasomal degradation. Degradation is required for termination of NF-kappa-B response.
Monomethylated at Lys-310 by SETD6. Monomethylation at Lys-310 is recognized by the ANK repeats of EHMT1 and promotes the formation of repressed chromatin at target genes, leading to down-regulation of NF-kappa-B transcription factor activity. Phosphorylation at Ser-311 disrupts the interaction with EHMT1 without preventing monomethylation at Lys-310 and relieves the repression of target genes.
Phosphorylation at Ser-311 disrupts the interaction with EHMT1 and promotes transcription factor activity (By similarity). Phosphorylation on Ser-536 stimulates acetylation on Lys-310 and interaction with CBP; the phosphorylated and acetylated forms show enhanced transcriptional activity.
Reversibly acetylated; the acetylation seems to be mediated by CBP, the deacetylation by HDAC3. Acetylation at Lys-122 enhances DNA binding and impairs association with NFKBIA. Acetylation at Lys-310 is required for full transcriptional activity in the absence of effects on DNA binding and NFKBIA association. Acetylation can also lower DNA-binding and results in nuclear export. Interaction with BRMS1 promotes deacetylation of 'Lys-310'.
- Cellular localizationNucleus. Cytoplasm. Nuclear, but also found in the cytoplasm in an inactive form complexed to an inhibitor (I-kappa-B). Colocalized with RELA in the nucleus upon TNF-alpha induction.
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Anti-NFkB p65 antibody [E379] images
Anti-NFkB p65 antibody [E379] (ab32536) at 1/100000 dilution + HeLa cell lysate
Predicted band size : 65 kDa
Observed band size : 65 kDa
Immunohistochemical analysis of paraffin-embedded human Breast carcinoma using anti-NF-kB p65 Rabbit Monoclonal Antibody
Immunofluorescent staining of HeLa cells using anti-NF-kB p65 Rabbit Monoclonal Antibody (ab32536)
Overlay histogram showing MCF-7 cells stained with ab32536 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32536, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in MCF-7 cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
Immunohistochemical analysis of colon sections from mice, staining NFkB p65 with ab32536.
Antigen retrieval was performed by microwave heating in citrate buffer, pH 6. Sections were incubated overnight with primary antibody (1/250) and staining was detected using ab80437 EXPOSE Rabbit specific HRP/DAB detection IHC kit.
References for Anti-NFkB p65 antibody [E379] (ab32536)
This product has been referenced in:
- Bel S et al. Loss of TMF/ARA160 protein renders colonic mucus refractory to bacterial colonization and diminishes intestinal susceptibility to acute colitis. J Biol Chem 287:25631-9 (2012). IHC-P ; Mouse . Read more (PubMed: 22553199) »
- Liu Y et al. IL-17 contributes to cardiac fibrosis following experimental autoimmune myocarditis by a PKCß/Erk1/2/NF-?B-dependent signaling pathway. Int Immunol 24:605-12 (2012). Read more (PubMed: 22531062) »