Influence of the protein staining in the fast ultrasonic sample treatment for protein identification through peptide mass fingerprint and matrix-assisted laser desorption ionization time of flight mass spectrometry

Journal of Proteome Research
M GalesioJ L Capelo

Abstract

The influence of the protein staining used to visualize protein bands, after in-gel protein separation, for the correct identification of proteins by peptide mass fingerprint (PMF) after application of the ultrasonic in-gel protein protocol was studied. Coomassie brilliant blue and silver nitrate, both visible stains, and the fluorescent dyes Sypro Red and Sypro Orange were evaluated. Results obtained after comparison with the overnight in-gel protocol showed that good results, in terms of protein sequence coverage and number of peptides matched, can be obtained with anyone of the four stains studied. Two minutes of enzymatic digestion time was enough for proteins stained with coomassie blue, while 4 min was necessary when silver or Sypro stainings were employed in order to reach equivalent results to those obtained for the overnigh in-gel protein protocol. For the silver nitrate stain, the concentration of silver present in the staining solution must be 0.09% (w/v) to minimize background in the MALDI mass spectra.

References

Mar 1, 1996·Analytical Chemistry·A ShevchenkoM Mann
May 3, 2001·Electrophoresis·W M LauberJ P Malone
Mar 30, 2004·Proteomics·Sophie RichertThierry Rabilloud
Oct 11, 2005·Journal of Proteome Research·D López-FerrerJ Vázquez
Oct 14, 2005·Bioscience Reports·Reiner Westermeier, Rita Marouga
Sep 23, 2006·Proteomics·Ingrid MillerElisabetta Gianazza
Feb 3, 2007·Journal of Proteome Research·R Rial-OteroJ L Capelo

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Citations

Mar 13, 2014·Talanta·J E AraújoJ L Capelo
Sep 14, 2012·Proteome Science·Carolina Fernández-CostaFrancisco J Blanco
Mar 1, 2011·Analytical Biochemistry·Seongjae ShinJeongkwon Kim
Oct 15, 2011·Electrophoresis·Carolina FernándezJosé-Luis Capelo-Martínez
Oct 27, 2016·ACS Applied Materials & Interfaces·Yoonjee C ParkJoyce Y Wong

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