Thermal shift assay

The ease of assay development and conduction of the actual melting experiment, the capability for high-throughput using microtiter-based readers, and the low requirements of reagents and protein consumption have recently resulted in 

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Fluorescence and fluorescence-based thermal shift assay 359... 

Pantoliano MW, Petrella EC, Kwasnoski JD, Lobanov VS, Myslik J, GrafE, Carver T, Asel E, Springer BA, Lane P, Salemme FR High-Density miniaturized thermal shift assays as a general strategy for drug discovery. / Biomol Screen 2001, 6, 429-440. 

MW Pantoliano, AW Rhind, FR Salemme. Microplate thermal shift assay for ligand development and multivariable protein chemistry optimization. U.S. Patent 6,020,141, 3-Dimensional Pharmaceuticals, 2000. 

Stabilization of the protein target by buffer additives or complex formation is a powerful way to enhance crystal-lizabihty. Biophysical techniques such as thermal-shift assays and NMR can be used for the identification of suitable hgands or additives. 

Lo M-C, Aulabaugh A, Jin G, Cowling R, Bard J, Malamas M, Ellestad G (2004) Evaluation of fluorescence-based thermal shift assays for hit identification in drug discovery. Anal Biochem 332 153-159... 

Independent confirmation of primary hits is performed by a variety of reliable secondary assays. We use a number of direct binding assays such as biolayer interferometry (BLI) or (for strongly binding inhibitors) ITC as well as indirect binding assays such as thermal shift assays for hit validation. 

Molina DM et al (2013) Monitoring drug target engagement in cells and tissues using the cellular thermal shift assay. Science 341 84-87... 

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