The PI3-Kinase Activity ELISA: Pico is a competitive ELISA that measures Class I Phosphoinositide 3-kinases (PI3-K) activity directly through its reaction product, PtdIns(3,4,5)P3 (PIP3). This assay can be used for in vitro determination of PI3-K activity in cancer tissues and cell lines or compound screening for drug discovery
Assay sensitivity: This assay can measure as low as 0.22 pmol PIP3 and detect activity from as little as 31 ng/mL PI3-K enzyme
Assay Incubation time: 2.5 hr
Sample Type: Purified and/ or Immunoprecipitated (IP) PI3-K Enzyme (see IP protocol at the end of the K-1000s TDS)
PI3-K reactions are run with the substrate PI(4,5)P2 (PIP2). The enzyme reactions, PIP3 standards, and controls are then mixed and incubated with PIP3 binding protein that is highly specific and sensitive to PIP3. This mixture is then transferred to a PIP3-coated microplate for competitive binding. Afterwards, a peroxidase-linked secondary detector and colorimetric detection is used to detect the amount of PIP3 produced by PI3-K. The amount of enzyme product (PIP3) produced is determined by comparing the enzyme reactions with a PIP3 standard curve.
PI3-K’s are ubiquitously expressed lipid kinases which phosphorylate PIP2 at the 3′-hydroxyl of the inositol ring producing PIP3. PIP3 has shown to be an important lipid second messenger with key roles in fundamental cellular responses such as proliferation and survival. Due to these interesting biological roles, PI3-K and PIP3 have become attractive targets for research and drug development in many diseases including inflammation and cancer.
Echelon has multiple formats available for Class III PI3-K. Please see this PI3-K Assay Comparison Guide.
Notice to Purchaser: Echelon Biosciences products are sold for research and development purposes only and are not to be incorporated into products for resale without written permission from Echelon Biosciences. This kit and all non-radioactive, competitive assays for determining phosphoinositide-3-kinase (PI3-K) activity is covered by Echelon Biosciences Inc. U.S. Patent 7,067,269. The purchase of this product includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser’s own internal research. For inquiries email busdev@echelon-inc.com.
Product Keywords: PI3-Kinase, PI3K, Phosphatidylinositol 3-kinases, PI 3-kinases, PI3Ks, K-1000