Anti-NF-κB p65 (Rel A) Rabbit Polyclonal Antibody
Catalog No: KC-5N01 Size: 100 µl (10 Western mini-blots) |
Applications |
Species Cross-Reactivity |
Molecular Wt. |
Source |
W, IP |
H, M, R |
65 kDa |
Rabbit |
Western blot anlaysis of extracts from HeLa cells, untreated or TNF-α treated for the indicated times, using Phospho-NF-κB p65 (Ser536) Antibody (top) or NF-κB p65 Antibody (bottom). |
Background:
The transcriptional nuclear factor kappa B (NF-κB) plays a pivotal role in inflammatory and immune responses. NF-κB was originally identified as a heterodimeric DNA binding protein complex consisting of p65 (RelA) and p50 (NF-κB1) subunits. Other identified subunits include p52 (NF-κB2), c-Rel, and RelB. The p65, cRel, and RelB subunits are responsible for transactivation. The p50 and p52 subunits possess DNA binding activity but limited ability to transactivate. p52 has been reported to form transcriptionally active heterodimers with the NF-κB subunit p65, similar to p50/p65 heterodimers. NF-κB is composed of a heterodimer of p65 and p50 subunits in most cell types and is sequestered in the cytoplasm by its inhibitory proteins, the IκBs. IκB-α binds to the p65 subunit (nuclear localization sequence, NLS) and IκB-α binding masks the NF-κB nuclear localization sequence (NLS), preventing NF-κB nuclear localization and DNA binding. NF-κB activating agents can induce the phosphorylation of IκBs; the phosphorylation targets IκBs for rapid degradation through an ubiquitin-proteasome pathway, thereby releasing NF-κB to enter the nucleus for gene expression. It has been shown that the p65 NF-κB subunit is also phosphorylated during the phosphorylation and degradation of IκBs; the phosphorylation site is Ser536 in the C-terminal transactivation domain. Phosphorylation of NF-κB p65 at serine 276 allows for increased intrection with the transcriptional coactiveator p300/CBP to further enhance the transcriptional activeity of this transcription factor.
Specificity/Sensitivity:
NF-κB p65 Antibody detects total endogenous levels of p65, a rel A gene product.
Source/Purification:
Polyclonal antibodies are produced by immunizing rabbits with a synthetic peptide (KLH coupled) corresponding to amino acids near c-terminus human p65.
Storage and Stability:
Each vial contains 100ul affinity purified rabbit anti- NFκB p65 (Rel A) antibody in a special stabilizer. Real time studies that this special stabilizer helped retain most of antibody activity for 24 months while store at 4℃.
Recommended Antibody Dilutions:
Western blotting |
1:1000 |
Immunoprecipitation |
1:250 |