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.
Supplied Product:
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.
Storage Instruction:
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.
Cytokine detection by antibody-based proximity ligation. Gullberg M, Gustafsdottir SM, Schallmeiner E, Jarvius J, Bjarnegard M, Betsholtz C, Landegren U, and Fredriksson S. Proc Natl Acad Sci U S A. 2004 Jun 1;101(22):8420-4. Epub 2004 May 21.
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