PI3 kinase p110α(E545K)/p85α Recombinant

Catalog #

40640

$455 *

Size: 20 µg

Qty

Bulk/Custom

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Description

Complex of human catalytic subunit p110α (phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha), and human regulatory subunit p85α (phosphoinositide-3-kinase regulatory subunit 1). The two proteins were co-expressed and affinity purified. Recombinant human p110α, encompassing full-length amino acids 2-1068(end). This construct contains an N-terminal Flag-tag and mutation of interest (E545K). Recombinant human p85α, full length encompassing amino acids 1-724(end). This construct does not contain a tag.

This product has been cited 2 times.

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Synonyms

PIK3CA, p110alpha, Phosphoinositide-3-Kinase, Catalytic, Alpha, PIK3R1, Phosphoinositide-3-Kinase, Regulatory Subunit 1 (Alpha), GRB1, p85alpha

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Product Data Gallery

10% SDS-PAGE Coomassie Staining

Specific Activity

Product Info

Storage and Usage

Citations2

Species

Human

Host Species/Expression System

Sf9 insect cells

Purity

≥87%

Format

Aqueous buffer solution

Formulation

25 mM Tris-HCl, pH 8.0, 69 mM NaCl, 1.35 mM KCl,0.025% Tween-20, 50% Glycerol and 3 mM DTT

p110α(E545K): 125 kDa; p85α: 84 kDa

Amino Acids

p110α(E545K): 2-1068(full length); p85α: 1-724(full length)

Specific Activity

1,200 pmol/min/μg

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.

Genbank #

p110α(E545K): NM_006218; p85α: NM181523

UniProt #

p110α(E545K): P42336; p85α: P27986

Tag(s)

p110α(E545K): N-terminal FLAG-tag

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)

Storage/Stability

At least 6 months at –80°C.

Assay Conditions

A 25 μl kinase reaction is conducted in 10 mM Tris-HCl (pH 7.5), 50 mM NaCl, 5 mM MgCl2, 100 μM ATP, 10 μg mix of PS and Phosphoinositol-4,5-Bisphosphate, and 50 ng enzyme for 15 minutes at 30°C. Conversion of ATP to ADP is detected using PI3K ADP-Glo® Luminescent Kinase Assay (Promega Corp., Madison, WI).

Instructions for Use

Thaw on ice and gently mix prior to use. DO NOT VORTEX. Perform a quick spin before opening. Aliquot into small volumes and flash freeze for long term storage. Avoid multiple freeze/thaw cycles.

Applications

Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.

Shipping Temperature

-80°C

Warnings

Avoid freeze/thaw cycles

Scientific Category

Kinase/Lipids

Regulatory Status

Research Use Only

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Datasheets/Protocols

40640 Lot 120409

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PI3 kinase p110α(E545K)/p85α Recombinant

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