ALEXIS Biochemicals: Phosphoinositide 3-kinase p110γ (human) (recombinant)

ALX-201-055

Revised 17-Apr-07

Phosphoinositide 3-kinase p110γ (human) (recombinant)
SYNONYMS	PI(3)K p110γ (human) (recombinant)
PRODUCT LINE	Signal Transduction
PRODUCT CATEGORY	Phosphoinositide 3-kinase [PI(3)K] / Related Products

Ordering Information

Product Numbers:	Format:	Size:	Unit Price:	Quantity:	Add To Cart
ALX-201-055-C010		10 µg	330.00 USD

Product Specification

EC:	2.7.1.137
SOURCE/HOST:	Produced in Sf9 cells. Full length PI(3)K p110γ (human phosphoinositide-3-kinase p110γ) is fused at the N-terminus to a His-tag.
CONCENTRATION:	1mg/ml
PURITY:	≥95% (SDS-PAGE)
FORMULATION:	Liquid. In 10mM HEPES, pH 7.5, containing 100mM sodium chloride, 0.5mM magnesium chloride and 50% glycerol.
SPECIFIC ACTIVITY:	~4.5nmol/mg/min using phosphatidylinositol as the substrate (lipid kinase activity test).
SHIPPING:	SHIPPED ON DRY ICE
LONG TERM STORAGE:	-20°C

Product Specific Literature References

Cloning and characterization of a G protein-activated human phosphoinositide-3 kinase: B. Stoyanov, et al.; Science 269, 690 (1995) Abstract
Linkage of G protein-coupled receptors to the MAPK signaling pathway through PI 3-kinase gamma: M. Lopez-Ilasaca, et al.; Science 275, 394 (1997) Abstract
Lipid kinase and protein kinase activities of G-protein-coupled phosphoinositide 3-kinase gamma: structure-activity analysis and interactions with wortmannin: S. Stoyanova, et al.; Biochem. J. 324, 489 (1997) Abstract; Full Text
Gbetagamma stimulates phosphoinositide 3-kinase-gamma by direct interaction with two domains of the catalytic p110 subuni: D. Leopoldt, et al.; J. Biol. Chem. 273, 7024 (1998) Abstract; Full Text

General Information

BACKGROUND/TECHNICAL INFORMATION

Swiss-Prot link P48736: PI(3)K p110γ (human)

Lipid kinase activity test:
Kinase reaction buffer: 20mM TRIS-HCl, pH 7.4; 4mM MgCl₂; 100mM sodium chloride.
- Reaction mixture in total volume of 100ml was:
1x reaction buffer (final concentration), enzyme (4 reactions with 1, 2, 3 or 4µg were performed), 10µg phosphatidylinositol, 20µM cold ATP and 10µCi ³²P-ATP.
- Reaction was started by addition of the ³²P-ATP and incubate for 10 minutes at 37°C.
- Reaction was stopped by adding 150µl 1M HCl.
- Lipids were extracted with 400µl chloroform:methanol (1:1) by vortexing.
- Extracts were washed three times with 200µl 1M HCl.
- Incorporation of ³²P in the extracted lipids was measured by mixing extract with 3ml scintillation cocktail (ROTH) and counting in a liquid scintillation analyzer.
- To determine specific activity of PI3K p110γ the assay was performed under the same conditions stated above by using 10µCi ³²P-ATP with a value of 10'000 units.
- Average value of measurements of the lipid extracts of the performed 4 reactions was 227 units for 1µg. Dividing 10’000 by 227 a factor of 44 times less for the incorporation was determined.
-Total ATP (20µM) was divided by 44 giving 0.454µM for 10 minutes reaction time or 45.4M/min/µg. Recalculation of this value for a 1 liter solution results in 4.5 nmoles/min/mg.

Further Categories Containing This Product:

Recombinant Proteins / Fusion Proteins • Enzymes

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