TNF Induces phosphorylation of RELA in KBM-5 cells. Nuclear protein lysates prepared after 0, 5, 10, 15, 30 and 60 minutes of 0.1 nM TNF treatment of KBM-5 cells shows inducible phosphorylation using RELA (phospho S529) polyclonal antibody (Cat # PAB10337). Anti-beta actin staining confirms loading of equivalent amounts of protein. HRP conjugated Gt-anti-Rabbit IgG was used to develop the blot using a chemiluminescent detection method. Data contributed by Aggarwal BB, personal communication.
Western Blot (Cell lysate)
Anti-pS529 shows phospho RELA staining in carcinoma cells. Immunoblot of total protein lysates from various human head and neck tumors shows phospho RELA staining in tumor cell lines using p65. Lanes 1 - 6 contain protein lysates from human squamal carcinoma cell lines. Lane 7 is a protein lysate from a primary culture of human keratinocytes. Lane 8 contains protein lysate from ATCC SCC9 cells (also a head and neck squamal carcinoma). Lane 9 contains lysate from EGF-induced human derived A-431 cells. Lane 10 (not shown) contains a molecular weight standard. Concurrent staining with anti-beta microtubulin (not shown) was used to confirmequal protein loading in all lanes. HRP conjugated Gt-anti-Rabbit IgG was used to develop the blot using a chemiluminescent detection method. Data contributed by Yu, M., NIH, personal communication.
NFKB1 (MIM 164011) or NFKB2 (MIM 164012) is bound to REL (MIM 164910), RELA, or RELB (MIM 604758) to form the NFKB complex. The p50 (NFKB1)/p65 (RELA) heterodimer is the most abundant form of NFKB. The NFKB complex is inhibited by I-kappa-B proteins (NFKBIA, MIM 164008 or NFKBIB, MIM 604495), which inactivate NFKB by trapping it in the cytoplasm. Phosphorylation of serine residues on the I-kappa-B proteins by kinases (IKBKA, MIM 600664, or IKBKB, MIM 603258) marks them for destruction via the ubiquitination pathway, thereby allowing activation of the NFKB complex. Activated NFKB complex translocates into the nucleus and binds DNA at kappa-B-binding motifs such as 5-prime GGGRNNYYCC 3-prime or 5-prime HGGARNYYCC 3-prime (where H is A, C, or T; R is an A or G purine; and Y is a C or T pyrimidine).[supplied by OMIM
nuclear factor of kappa light polypeptide gene enhancer in B-cells 3,v-rel avian reticuloendotheliosis viral oncogene homolog A (nuclear factor of kappa light polypeptide gene enhancer in B-cells 3 (p65)),v-rel reticuloendotheliosis viral oncogene homolog