Complex of N-terminal FLAG-tagged recombinant full-length human p110alpha (GenBank Accession No. NM_006218), and recombinant full length, human p85alpha (no tag) (GenBank Accession No. NM_181523). Coexpressed in a Baculovirus-infected Sf9 cell expression system. p110alpha MW=125 kDa, p85alpha MW=84 kDa.
Host cell line: Sf9 cells
Specific activity: 1500 pmol/min/ug.
Unit definition: A unit of p110alpha/p85alpha kinase activity is defined as the amount of enzyme required to produce 1 pmol of Phosphoinositol-3, 4,5-Trisphosphate/min at 37°C.
Immunogen Region
full length
Tag
N-terminal FLAG-tag(p110alpha), no tag on p85alpha
Formulation
25 mM Tris, pH 8.0, 69 mM NaCl, 1.35 mM KCl, 0.025% Tween-20, 3 mM DTT, and 50% glycerol.
Species
Human
Application
Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling
Notes
A 25 ul kinase reaction is conducted in a buffer containing 40 mM Tris-HCl (pH 7.4), 20 mM MgCl2, 100 uM ATP, 200 uM PI:PS substrate (BPS catalog #40560) and 110alpha/p85alpha enzyme for 15 minutes at 30°C. ATP reduction is detected using ADP-Glo Kinase Assay Platform (Promega Corporation, Madison, WI).