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Heat-mediated antigen retrieval is recommended prior to staining, using a 10mM citrate buffer, pH 6.0, for 10-20 minutes followed by cooling at room temperature for 20 min. Following antigen retrieval, incubate samples with primary antibody for 10 min at room temperature. A suggested positive control is placenta or breast carcinoma.
Phosphatidylinositol 3-kinase (PIK3) is a signaling protein that phosphorylates the inositol ring of phosphatidylinositol at the 3-prime position. Signaling is initiated when upstream tyrosine kinase receptors or G protein coupled receptors are bound by their ligands. It binds to activated (phosphorylated) protein-Tyr kinases, through its SH2 domain, and acts as an adapter, mediating the association of the p110 catalytic unit to the plasma membrane. Phosphatidylinositol 3-kinase plays an important role in the metabolic actions of insulin, and a mutation in this gene has been associated with insulin resistance. During insulin stimulation, it also binds to IRS-1. Mutations in the gene can result in Agammaglobulinemia 7, SHORT syndrome, or Immunodeficiency 36.
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