Representative image of Proximity Ligation Assay of protein phosphorylation. HeLa cells were stained with dual recognition antibody pair set, rabbit polyclonal antibody 1:1200 and mouse monoclonal antibody 1:50. Each red dot represents one single phosphorylated protein. The images were analyzed using an optimized freeware (BlobFinder) download from The Centre for Image Analysis at Uppsala University.
Antibody pair set content: 1. Phospho-RELA T254 rabbit polyclonal antibody (20 ul) In PBS, 150 mM NaCl, pH 7.4 (0.02% sodium azide, 50% glycerol) 2. RELA mouse monoclonal antibody (40 ug) In 1x PBS, pH 7.2 *Reagents are sufficient for at least 30-50 assays using recommended protocols.
Store reagents of the antibody pair set at -20°C or lower. Please aliquot to avoid repeated freeze thaw cycle. Reagents should be returned to -20°C storage immediately after use.
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