Please use this identifier to cite or link to this item: http://hdl.handle.net/2381/14782
Title: Digestion of native proteins for proteomics using a thermocycler.
Authors: Turapov, OA
Mukamolova, GV
Bottrill, AR
Pangburn, MK
First Published: 1-Aug-2008
Citation: ANAL CHEM, 2008, 80 (15), pp. 6093-6099
Abstract: Efficient protein digestion is a critical step for successful mass spectrometry analysis. Here we describe simultaneous tryptic digestion and gradual unfolding of native proteins by application of a temperature gradient using a single cycle of 5 min or less in a PCR thermocycler. Chemicals typically used for chromatographic techniques did not affect the digestion efficiency. Tryptic digestion was performed in a small volume (3 microL) with 1.5 microg of trypsin without denaturing agents. This rapid procedure yielded more peptides than conventional methods utilizing chemical denaturation for 18 proteins out of 20. Samples were directly spotted on the MALDI-TOF target plate, without additional purification, thus reducing losses on reversed-phase resins.
DOI Link: 10.1021/ac702527b
eISSN: 1520-6882
Links: http://hdl.handle.net/2381/14782
Type: Journal Article
Appears in Collections:Published Articles, Dept. of Infection, Immunity and Inflammation

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