Products from BPS Bioscience require a minimum order value above 400€
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 p110α(E545K)/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 FLAG-tagged recombinant full-length human p110α (GenBank Accession No. NM_006218), with E545K mutation, and recombinant full length, human p85α (no tag, GenBank Accession No. NM_181523). Co-expressed in Sf9 cells in a Baculovirus expression system. p110α(E545K) MW=125 kDa, p85α MW=84 kDa.
Format: Aqueous buffer solution
Formulation: 25 mM Tris-HCl, pH 8.0, 69 mM NaCl, 1.35 mM KCl, 0.03% Tween-20, 3 mM DTT, 50% glycerol.
Genbank: NM_006218 (p110α), NM_181523 (p85α)
Host Cell Line: Sf9 cells
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal FLAG-tag(p110α), no tag on p85α
Uniprot: p110α: P42336; p85α: P27986
Unit Definition: A unit of p110α(E545K)/p85α kinase activity is defined as the amount of enzyme required to produce 1 pmol of Phosphoinositol-3, 4,5-Trisphosphate/min at 37°C.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Engelman, J.A., et al. Nature Med. 14:1351-1356 (2008)
2. Hafner, C., et al., Proc. Natl. Acad. Sci. USA. 104(33):13450-13454 (2007)
3. Serra, V., et al., Cancer Res.68(19):8022-8030 (2008)