Methods in molecular biology (Clifton, N.J.), 2012, Vol.803, p.141
2012
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Details
- Autor(en) / Beteiligte
- Titel
- Determination of kinase inhibitor potencies in cell extracts by competition binding assays and isobaric mass tags
- Ist Teil von
- Methods in molecular biology (Clifton, N.J.), 2012, Vol.803, p.141
- Ort / Verlag
- United States
- Erscheinungsjahr
- 2012
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- Chemical proteomics offers a unique approach for target identification of small molecule inhibitors directly from cell extracts, thus enabling characterization of target proteins under close to physiological conditions. Here, we describe a competition binding procedure that is based on affinity enrichment of potential target proteins on a probe matrix in the presence of increasing amounts of free test compound in solution. Reduced binding of target proteins to the probe matrix as a function of test compound concentration can be measured and thus, enables calculation of IC(50) values. The method employs quantitative mass spectrometry using isobaric mass tags which enables determination of potency for a large number of target proteins in a single analysis.
- Sprache
- Englisch
- Identifikatoren
- eISSN: 1940-6029DOI: 10.1007/978-1-61779-364-6_10Titel-ID: cdi_pubmed_primary_22065223
- Format
- –
- Schlagworte
- Binding, Competitive - drug effects, Biological Assay - methods, Cell Extracts - chemistry, Cell Fractionation, Chromatography, Affinity, Chromatography, Liquid, Inhibitory Concentration 50, Mass Spectrometry, Molecular Weight, Protein Kinase Inhibitors - pharmacology, Proteomics, Staining and Labeling, Statistics as Topic, Staurosporine - pharmacology, Trypsin