PIP Arrays are membranes pre-spotted with a concentration gradient of all eight phosphoinositides. PIP Arrays are used in a simple protein-lipid overlay assay to help determine the specificity and sensitivity of your protein towards one or more of these lipids.
Briefly, once the membrane is blocked, it is incubated with your protein of interest. The membrane is then washed and incubated with an HRP labeled antibody against the protein or your proteins affinity tag. The membrane is washed again before incubation with ECL or a precipitating TMB detection reagent. This can then be visualized with the appropriate equipment based on the detection reagent used.
PIP Arrays are a 4 x5 cm hydrophobic membrane with 100, 50, 25, 12.5, 6.25, 3.13, and 1.56 pmol of the following lipids: Phosphatidylinositol (PtdIns), Phosphatidylinositol (3)-phosphate (PtdIns(3)P), Phosphatidylinositol (4)-phosphate (PtdIns(4)P), Phosphatidylinositol (5)-phosphate (PtdIns(5)P), Phosphatidylinositol (3,4)-bisphosphate (PtdIns(3,4)P2 ), Phosphatidylinositol (3,5)-bisphosphate (PtdIns(3,5)P2), Phosphatidylinositol (4,5)-bisphosphate (PtdIns(4,5)P2), Phosphatidylinositol (3,4,5)-trisphosphate (PtdIns(3,4,5)P3).
Each 5-pack of PIP Arrays includes a free vial of the PI(4,5)P2 specific binding protein for use as a positive control. For a list other phosphoinositide binding proteins, click here.