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-AKT1 Y315 rabbit polyclonal antibody (20 ul) In PBS (without Mg2+ and Ca2+), 150 mM NaCl, pH 7.4 (0.02% sodium azide, 50% glycerol) 2. AKT1 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.
The serine-threonine protein kinase encoded by the AKT1 gene is catalytically inactive in serum-starved primary and immortalized fibroblasts. AKT1 and the related AKT2 are activated by platelet-derived growth factor. The activation is rapid and specific, and it is abrogated by mutations in the pleckstrin homology domain of AKT1. It was shown that the activation occurs through phosphatidylinositol 3-kinase. In the developing nervous system AKT is a critical mediator of growth factor-induced neuronal survival. Survival factors can suppress apoptosis in a transcription-independent manner by activating the serine/threonine kinase AKT1, which then phosphorylates and inactivates components of the apoptotic machinery. Multiple alternatively spliced transcript variants have been found for this gene. [provided by RefSeq