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Encompassing Amino Acids: full length
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: A 25 µl kinase reaction is conducted in a buffer containing 40 mM Tris-HCl (pH 7.4), 20 mM MgCl2, 100 uM ATP, 200 µM PI:PS substrate (BPS catalog #40560) and p110δ/p85α enzyme for 15 minutes at 30°C. ATP reduction is detected using ADP-Glo™ Kinase Assay Platform (Promega Corporation, Madison, WI).
Description: Complex of N-terminal GST-tagged recombinant full-length human p110δ (GenBank Accession No. NM_005026.2), and recombinant full length, human p85α, GenBank Accession No. NM_181523). Coexpressed in a Baculovirus infected Sf9 cell expression system. p110δ MW=146 kDa, p85α MW=84 kDa.
Format: Aqueous buffer solution
Formulation: 25 mM Tris, pH 8.0, 69 mM NaCl, 1.35 mM KCl, 0.03% Tween-20, 3 mM DTT, 50% glycerol.
Genbank: NM_005026.2 (p110δ), NM_181523 (p85α)
Host Cell Line: Sf9 cells
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal GST-tag (p110δ), no tag on p85α
Uniprot: p110δ: O00329; p85α: P27986
Unit Definition: A unit of p110δ/p85α kinase activity is defined as the amount of enzyme required to produce 1 pmol of Phosphoinositol-3,4,5-Trisphosphate/min at 30°C.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Vanhaesebroeck et al. (1997) Proc. Natl. Acad. Sci. U.S.A. 94:4330.
2. Vanhaesebroeck et al. (1999) EMBO J. 18:1292.
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