The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/250. Detects a band of approximately 180 kDa (predicted molecular weight: 131 kDa).
May mediate the control of various cellular processes by insulin. When phosphorylated by the insulin receptor binds specifically to various cellular proteins containing SH2 domains such as phosphatidylinositol 3-kinase p85 subunit or GRB2. Activates phosphatidylinositol 3-kinase when bound to the regulatory p85 subunit.
Polymorphisms in IRS1 may be involved in the etiology of non-insulin-dependent diabetes mellitus (NIDDM) [MIM:125853].
Serine phosphorylation of IRS1 is a mechanism for insulin resistance. Ser-312 phosphorylation inhibits insulin action through disruption of IRS1 interaction with the insulin receptor. Phosphorylation of Tyr-896 is required for GRB2-binding.
The 180kDa band is believed to correspond to phospho S307 IRS1.
Blocking with the immunising peptide removes the 180kDa band (data not shown). A 4-20% Tris-Glycine gel was used, with blocking with 5% goat serum and 0.5% BLOTTO in PBS.