AffiASSAY® Phosphatidylinositol Assay Kit (Fluorometric)

Specifications

Description

Phosphoinositides are critical second messengers in a multitude of cellular processes. The most fundamental phosphoinositide, phosphatidylinositol (PI), can be phosphorylated at multiple sites around its inositol ring, leading to downstream activation. For instance, phosphatidylinositol 4-[mono]phosphate is involved in membrane trafficking, while cleavage of phosphatidylinositol triphosphate yields inositol triphosphate, a molecule involved in calcium signaling. Signaling is conveyed in many signaling proteins (including Protein Kinase B/Akt) via their domain referred to as a Pleckstrin homology, or PH domain, which is capable of binding to specific phosphoinositides. Biosynthesis of PI involves the action of phosphatidylinositol synthase on cytidine diacylglycerol and myo-inositol. The critical signal transducer enzymes, Phosphatidylinositolinositol-3-kinases phosphorylate PI at the 3 position hydroxyl of inositol, leading to modulation in pathways such as cell growth, survival, and motility. PI is typically at concentrations in the low- to mid-micromolar range in human serum and plasma, and is capable of activating second messengers such as Ca2+ and cyclic AMP. It is through these pathways that PI has been found to affect such biological events as the pro-inflammatory response and intestinal uptake. BioVision’s Phosphatidylinositol Assay Kit utilizes specific enzymes to generate an intermediate that then reacts with PicoProbeTM, yielding a signal that can be quantified fluorometrically, and is proportional to the amount of PI present in the sample. The assay is capable of detecting as little as 15 pmoles of phosphatidylinositol.