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Effect of staining reagent on peptide mass fingerprinting from in‐gel trypsin digestions: A comparison of SyproRuby™ and DeepPurple™
Author(s) -
Tannu Nilesh S.,
SanchezBrambila Gabriela,
Kirby Pam,
Andacht Tracy M.
Publication year - 2006
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200500740
Subject(s) - stain , staining , trypsin , chromatography , proteome , chemistry , peptide mass fingerprinting , peptide , reagent , microbiology and biotechnology , biochemistry , proteomics , biology , enzyme , genetics , gene
As the new fluorescent stains such as SyproRuby™ and DeepPurple™ are getting widespread recognition for proteome analyses by the traditional 2‐D gel method, it becomes important to test the feasibility of these stains with respect to staining reproducibility, protein quantitation, and compatibility of the stain with downstream MS. The binding of epicocconone, active ingredient of DeepPurple™, to one of the primary cleavage sites of trypsin (lysine residue) raises the possibility of incomplete cleavage and interference with PMF. However, the current study tests and concludes that the DeepPurple™ stain can result in increased peptide recovery compared to SyproRuby™ stain and can improve MS‐based identification of lower intensity proteins spots.