PIP3 Quantification TR-FRET Assay

PI 3-Kinases α, β, γ and δ (h) catalyse the phosphorylation of phosphatidylinositol 4,5-bisphosphate (PIP2) to phosphatidylinositol 3,4,5-trisphosphate (PIP3) in the presence of ATP and Mg2+. The PIP3 product is detected by displacement of biotin-PIP3 from an energy transfer complex consisting of Europium labelled anti-GST monoclonal antibody, a GSTtagged pleckstrin homology (PH) domain, biotinylated PIP3 and Streptavidin-Allophycocyanin (APC). Excitation of Europium in the complex results in an energy transfer to the APC and a fluorescent emission at 665 nm. The PIP3 product isolated from cell membranes displaces biotin-PIP3 from the complex resulting in a loss of energy transfer and thus a decrease in signal.

Research Category
Signaling

The PIP3 Quantification TR-FRET Assay kit provides an easy to use method for assaying PIP3 levels by detecting the displacement of biotin-PIP3 from an energy transfer complex.

This assay requires lipid extraction from cells. A protocol is included with this assay.

Used to detect/quantify: PI 3-Kinase

Kit capacity: 96-wells

Black 96-well plate

Detection Buffer

DM B

DM A

Stop B

PIP3 Standard

Lipid Recovery Buffer

1M DTT

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.