Pi3-Kinase Activity Alphascreen Assay

The PI3-Kinase Activity AlphaScreen® Assay is a sensitive and robust method for the quantification of PI(3,4,5)P3 (PIP3), the end product of Class I PI3K reactions. This high-throughput (384-well) assay can be used for in vitro determination of PI3K activity in cancer tissues and cell lines and compound screening for drug discovery.

Assay Range:0.46 NM to 1,000 nM

Sample Type:purified enzyme, inhibitor screen

Sample Volume:2.5 µL PI3-K inhibitor

Product Background:When PIP3 is generated via phosphorylation of PI(4,5)P2 (PIP2) by PI3-Kinase, the products of the enzymatic reaction compete with biotinylated I(1,3,4,5)P4 for the interaction with the PIP3 detector protein. The interaction of biotinylated-I(1,3,4,5)P4 and the PIP3 detector protein brings both Acceptor and Donor beads together, producing a cascade of chemical reactions and leading to the AlphaScreen® signal. In the absence of this interaction, proximity of the Donor and Acceptor beads is decreased, producing a loss of luminescent signal which is inversely related to enzyme activity. AlphaScreen® technology is based on the emission of light (520-620nm) by Acceptor beads activated by the proximity of Donor beads.

Echelon has multiple formats available for Class III PI3-K. Please see the PI3-K Assay Comparison Guide below for more information.

Things to consider:This product is intended for use with the Amplified Luminescent Proximity Homogenous Assay (AlphaScreen®) technology and requires the researcher to obtain donor and acceptor beads from the AlphaScreen® GST (Glutathione-S-Transferase) Detection Kit, which must be purchased separately from PerkinElmer Life Sciences. A microplate reader with AlphaScreen® capabilities is also required.

Remark：These protocols are for reference only. Solarbio does not independently validate these methods.

Note：

1. The products are all for scientific research use only. Do not use it for medical, clinical diagnosis or treatment, food and cosmetics, etc. Do not store them in ordinary residential areas.

2. For your safety and health, please wear laboratory clothes, disposable gloves and masks.

3. The experimental results may be affected by many factors, after-sale service is limited to the product itself and does not involve other compensation.