Quantitative measurement of phosphatidylinositol 3,4,5-trisphosphate.

Quantitative measurement of phosphatidylinositol 3,4,5-trisphosphate. Research Abstract Details 

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Quantitative measurement of phosphatidylinositol 3,4,5-trisphosphate. Abstract Text:

Guillou,Len R Stephens,Phillip T Hawkins, Guillou,Len R Stephens,Phillip T Hawkins,

The activation of class I phosphoinositide 3-kinases (PI(3)Ks) by cell surface receptors represents the initiation of a large and complex signaling network that couples many growth factors, antigens, and inflammatory stimuli to important cellular responses, such as cell growth, survival, and movement. The most direct measurement of class I PI(3)K activity in cells is the rate of production of its lipid product, phosphatidylinositol 3,4,5-trisphosphate [PtdIns(3,4,5)P(3)]. This chapter describes in detail two approaches used to estimate the levels of PtdIns(3,4,5)P(3) in cells. One approach uses radiotracer labeling of cells, lipid extraction, deacylation, and subsequent quantitation of phosphoinositides by anion-exchange high-performance liquid chromatography. The second approach uses a novel, nonradioactive assay in which the cellular lipids are extracted, phosphoinositides are enriched through binding to a neomycin matrix, dried onto a nitrocellulose membrane, and PtdIns(3,4,5)P(3) quantified by a protein-lipid overlay approach using a GRP(1) PH domain probe.

Quantitative measurement of phosphatidylinositol 3,4,5-trisphosphate. Publishing Authors By Initials

H Guillou,LR Stephens,PT Hawkins,H Guillou,LR Stephens,PT Hawkins,

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Quantitative measurement of phosphatidylinositol 3,4,5-trisphosphate. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Methods in enzymology

VOLUME: 434

Page Numbers: 117-30

Journal Abbreviation: Meth. Enzymol.

ISSN: 0076-6879

DAY: 23

MONTH: 10

YEAR: 2007

Quantitative measurement of phosphatidylinositol 3,4,5-trisphosphate. Information

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LANGUAGE: eng

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Grant and Affiliation Information for Quantitative measurement of phosphatidylinositol 3,4,5-trisphosphate.

AFFILIATION: The Inositide Laboratory, The Babraham Institute, Babraham Research Campus, Cambridge, United Kingdom.

Country: United States

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MEDLINETA: Methods Enzymol

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