Mass spectrometric identification of proteins from silver-stained polyacrylamide gel: a method for the removal of silver ions to enhance sensitivity.
Mass spectrometry is a powerful technique for the identification of proteins at nanogram quantities. However, some degree of sample preparation prior to mass spectrometry is required, and silver-stained protein gel samples are most problematic. Here we report our strategy to obtain peptide mass profiles from silver-stained protein gel samples from one- or two-dimensional gels by destaining prior to enzymatic digestion. This study demonstrates that by using the destaining method, the sensitivity and quality of mass spectra is increased for matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometric analysis, permitting more proteins to be identified by peptide mass database analysis.
Gharahdaghi
F
F
The Rockefeller University Protein/DNA Technology Center, New York, NY 10021, USA. gharahf@rockvax.rockefeller.edu
Weinberg
C R
CR
Meagher
D A
DA
Imai
B S
BS
Mische
S M
SM
D000180:Acrylic Resins; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D007477:Ions; D011506:Proteins / Q000032:analysis*; D012680:Sensitivity and Specificity; D016622:Silver Staining / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D000180:Acrylic Resins; D007477:Ions; D011506:Proteins; C016680:polyacrylamide gels