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 purified polyclonal 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-EIF4E S209 rabbit polyclonal antibody (20 ul) In PBS, 150 mM NaCl, pH 7.4 (0.02% sodium azide, 50% glycerol) 2. EIF4E mouse purified polyclonal 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.
All eukaryotic cellular mRNAs are blocked at their 5-prime ends with the 7-methylguanosine cap structure, m7GpppX (where X is any nucleotide). This structure is involved in several cellular processes including enhanced translational efficiency, splicing, mRNA stability, and RNA nuclear export. EIF4E is a eukaryotic translation initiation factor involved in directing ribosomes to the cap structure of mRNAs. It is a 24-kD polypeptide that exists as both a free form and as part of a multiprotein complex termed EIF4F. The EIF4E polypeptide is the rate-limiting component of the eukaryotic translation apparatus and is involved in the mRNA-ribosome binding step of eukaryotic protein synthesis. The other subunits of EIF4F are a 50-kD polypeptide, termed EIF4A (see MIM 601102), that possesses ATPase and RNA helicase activities, and a 220-kD polypeptide, EIF4G (MIM 600495) (Rychlik et al., 1987 [PubMed 3469651]).[supplied by OMIM