PurificationAntibodies were produced by immunizing rabbits with synthetic peptide and KLH conjugates. Antibodies were purified by affinity-chromatography using epitope-specific peptide.
ApplicationsWB IHC
Species ReactivityHu
SpecificityThe antibody detects endogenous level of total NFkB-p65 protein.
Immunogen TypePeptide-KLH
Immunogen DescriptionPeptide sequence around aa.527~531 (L-L-S-G-D) derived from Human NFkB-p65.
Target NameNFkB-p65
Alternative NamesNFKB3; RELA; TF65; Transcription factor p65; p65
FormulationSupplied at 1.0mg/mL in phosphate buffered saline (without Mg2+ and Ca2+), pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
StorageStore at -20°C for long term preservation (recommended). Store at 4°C for short term use.
Application Details
Predicted MW: 65kd
Western blotting: 1:500~1:1000
Immunohistochemistry: 1:50~1:100
Images
Western blot analysis of extracts from 293 and Hela cells using NFkB-p65(Ab-529) Antibody #AB21210.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using NFkB-p65(Ab-529) Antibody #AB21210(left) or the same antibody preincubated with blocking peptide(right).
Background
NF-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
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