PI3 Kinase Activity ELISA Kit

Sample

Cancer tissue, cell lines

Species Reactivity

N/A

Detection Method

cELISA

Intended Use

ELISA kit that measures class 1 PI3-kinase activity directly through its reaction product, PIP3. This assay can be used for in vitro PI3K activity in cancer tissue, cell lines, or compound screening for drug discovery.

Contents of Kit

1. PI(3,4,5)P3 Detection Plate. 1 plate
2. PI(4,5)P2 Substrate, diC8. 23 nmol
3. PI(3,4,5)P3 Standard, diC8. 0.9 nmol
4. PI(3,4,5)P3 Detector. 20 μL
5. Detection Buffer. 10 mL
6. Secondary Detector. 300 μL
7. 10× TBS-T Buffer. 20 mL
8. TMB Solution. 12 mL
9. 1 N H₂SO₄ Stop Solution. 8 mL
10. 5× KBZ Kinase Reaction Buffer. 4 mL
11. 100 mM EDTA Solution. 500 μL
12. Incubation Plate. 1 plate
13. Plate Sealers. 3 seals

Storage

Store kit at 4°C. Store prepared reagents as indicated in the protocol

General Description

The production of PI(3,4,5)P3 from PI(4,5)P2 by PI3-Kinases (PI3-K) is important in multiple cell signaling pathways. Typically, experiments to measure PI3-K activity have involved phosphorylation of a phosphoinositide substrate using ³²P, then extraction of radioactive products, and separation using thin-layer chromatography or HPLC. The assay plate method developed by Creative diagnostics allows the user to determine PI3-K activity, using either recombinant or immunoprecipitated enzyme, in a standard ELISA format, eliminating the need for radioactivity, and thin layer chromatography or HPLC.