Identifying competitive protein antagonists for F-actin with reverse-phase high-performance liquid chromatography

Jeffrey W. Brown; C. James McKnight

doi:10.1016/j.ab.2009.11.023

Identifying competitive protein antagonists for F-actin with reverse-phase high-performance liquid chromatography

Jeffrey W. Brown
, C. James McKnight

Research output: Contribution to journal › Article › peer-review

4 Scopus citations

Abstract

F-actin binding constants are traditionally determined by centrifugal cosedimentation with actin microfilaments, where bound protein is separated from actin with SDS-PAGE and quantitated using densitometry. Here, we demonstrate that UV quantitation of reverse-phase HPLC-separated proteins provides increased accuracy and sensitivity, can be fully automated, and allows one to perform F-actin competition assays on similar sized proteins.

Original language	English
Pages (from-to)	117-119
Number of pages	3
Journal	Analytical Biochemistry
Volume	398
Issue number	1
DOIs	https://doi.org/10.1016/j.ab.2009.11.023
State	Published - Mar 1 2010

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10.1016/j.ab.2009.11.023

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title = "Identifying competitive protein antagonists for F-actin with reverse-phase high-performance liquid chromatography",

abstract = "F-actin binding constants are traditionally determined by centrifugal cosedimentation with actin microfilaments, where bound protein is separated from actin with SDS-PAGE and quantitated using densitometry. Here, we demonstrate that UV quantitation of reverse-phase HPLC-separated proteins provides increased accuracy and sensitivity, can be fully automated, and allows one to perform F-actin competition assays on similar sized proteins.",

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AU - McKnight, C. James

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N2 - F-actin binding constants are traditionally determined by centrifugal cosedimentation with actin microfilaments, where bound protein is separated from actin with SDS-PAGE and quantitated using densitometry. Here, we demonstrate that UV quantitation of reverse-phase HPLC-separated proteins provides increased accuracy and sensitivity, can be fully automated, and allows one to perform F-actin competition assays on similar sized proteins.

AB - F-actin binding constants are traditionally determined by centrifugal cosedimentation with actin microfilaments, where bound protein is separated from actin with SDS-PAGE and quantitated using densitometry. Here, we demonstrate that UV quantitation of reverse-phase HPLC-separated proteins provides increased accuracy and sensitivity, can be fully automated, and allows one to perform F-actin competition assays on similar sized proteins.

UR - https://www.scopus.com/pages/publications/73549103099

U2 - 10.1016/j.ab.2009.11.023

DO - 10.1016/j.ab.2009.11.023

M3 - Article

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SN - 0003-2697

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SP - 117

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JO - Analytical Biochemistry

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IS - 1

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Identifying competitive protein antagonists for F-actin with reverse-phase high-performance liquid chromatography

Abstract

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