Anti-NFkB p65 (acetyl K310) antibody - ChIP Grade (ab52175)
Overview
- Product nameAnti-NFkB p65 (acetyl K310) antibody - ChIP GradeSee all NFkB p65 primary antibodies ...
- DescriptionRabbit polyclonal to NFkB p65 (acetyl K310) - ChIP Grade
- Specificityab52175 detects endogenous levels of NF-kB p65 only when acetylated at lysine 310.
- Tested applicationsWB, ELISA, ChIP, ICC/IF more details
- Species reactivityReacts with: Mouse, Rat, Human
- Immunogen
Synthetic acetylated peptide derived from human NF-kB p65 around the acetylation site of lysine 310 (T-F-KA-S-I).
- Positive controlExtracts from HeLa cells treated with TSA (40nM, 24hours).
Properties
- FormLiquid
- Storage instructionsStore at -20°C. Stable for 12 months at -20°C
- Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 50% Glycerol, PBS (without Mg2+ and Ca2+), 150mM Sodium chloride, pH 7.4 -
Concentration information loading... - PurityImmunogen affinity purified
- Purification notesab52175 was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific acetylated peptide. The antibody against non-acetylated peptide was removed by chromatography using non-acetylated peptide corresponding to the acetylation site.
- Clonality Polyclonal
- IsotypeIgG
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Research Areas
Applications
Our Abpromise guarantee covers the use of ab52175 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Notes |
|---|---|
| WB | WB: 1/500 - 1/1000. Detects a band of approximately 70 kDa (predicted molecular weight: 60 kDa). |
| ELISA | ELISA: 1/40000. |
| ChIP | ChIP: Use at an assay dependent concentration. |
| ICC/IF | ICC/IF: Use a concentration of 1 - 5 µg/ml. |
Target
- 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.
- Post-translational
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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Database links
- Entrez Gene: 5970 Human
- Entrez Gene: 19697 Mouse
- Entrez Gene: 309165 Rat
- Omim: 164014 Human
- SwissProt: Q04206 Human
- SwissProt: Q04207 Mouse
- Unigene: 502875 Human
- Unigene: 249966 Mouse
- Unigene: 19480 Rat
see all
Target information above from: UniProt accession
Q04206
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010)
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Alternative names
- Avian reticuloendotheliosis viral (v rel) oncogene homolog A antibodyMGC131774 antibodyNF kappa B p65delta3 antibody
- NFKB 3 antibodyNFKB3 antibodyNuclear Factor NF Kappa B p65 Subunit antibodyNuclear factor NF-kappa-B p65 subunit antibodyNuclear factor of kappa light polypeptide gene enhancer in B cells 3 antibodyNuclear Factor Of Kappa Light Polypeptide Gene Enhancer In B Cells antibodyNuclear factor of kappa light polypeptide gene enhancer in B-cells 3 antibodyOTTHUMP00000233473 antibodyOTTHUMP00000233474 antibodyOTTHUMP00000233475 antibodyOTTHUMP00000233476 antibodyOTTHUMP00000233900 antibodyp65 antibodyp65 NF kappaB antibodyp65 NFkB antibodyRELA antibodyTF65_HUMAN antibodyTranscription Factor p65 antibodyv rel avian reticuloendotheliosis viral oncogene homolog A (nuclear factor of kappa light polypeptide gene enhancer in B cells 3 (p65)) antibodyV Rel Avian Reticuloendotheliosis Viral Oncogene Homolog A antibodyv rel reticuloendotheliosis viral oncogene homolog A (avian) antibodyv rel reticuloendotheliosis viral oncogene homolog A antibodyV rel reticuloendotheliosis viral oncogene homolog A, nuclear factor of kappa light polypeptide gene enhancer in B cells 3, p65 antibody
see all
Anti-NFkB p65 (acetyl K310) antibody - ChIP Grade images
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Western blot - NFkB p65 (acetyl K310) antibody (ab52175)All lanes : Anti-NFkB p65 (acetyl K310) antibody - ChIP Grade (ab52175) at 1/500 dilution
Lane 1 : HeLa cell extract treated with TSA (40nM, 24hours) with immunising peptide
Lane 2 : HeLa cell extract treated with TSA (40nM, 24hours)
Secondary
Alkaline Phosphatase-conjugated Monoclonal Mouse Anti-Rabbit.
Predicted band size : 60 kDa
Observed band size : 70 kDa (why is the actual band size different from the predicted?) -
Immunocytochemistry/ Immunofluorescence-NFkB p65 (acetyl K310) antibody(ab52175)ICC/IF image of ab52175 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab52175, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. -
Western blot - Anti-NFkB p65 (acetyl K310) antibody (ab52175)
developed using the ECL technique
Predicted band size : 60 kDa
Exposure time : 30 secondsImage courtesy of Tom Hu by Abreview.
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ChIP - Anti-NFkB p65 (acetyl K310) antibody (ab52175)Image courtesy of Tom Hu by Abreview.ab52175 used undiluted in ChIP.Whole tissue lysate prepared from murine sciatic nerves.10 seconds cross-linking (X-ChIP) with paraformaldehyde.Detection step: Semiquantitative PCR.
References for Anti-NFkB p65 (acetyl K310) antibody - ChIP Grade (ab52175)
This product has been referenced in:
- Ziesché E et al. The coactivator role of histone deacetylase 3 in IL-1-signaling involves deacetylation of p65 NF-?B. Nucleic Acids Res 41:90-109 (2013). Read more (PubMed: 23087373) »
- Clavijo PE & Frauwirth KA Anergic CD8+ T Lymphocytes Have Impaired NF-?B Activation with Defects in p65 Phosphorylation and Acetylation. J Immunol 188:1213-21 (2012). WB ; Mouse . Read more (PubMed: 22205033) »
