Mass spectrometry compatibility of two-dimensional gel protein stains.
As proteomic technology evolves, protein staining sensitivity is constantly being improved, enabling researchers to better visualize the proteome of their system. The current challenge is to balance the limits of detection of protein visualization with those of the mass spectrometric methods. In this report, mass spectra generated from human serum or rat liver proteins stained with either colloidal Coomassie blue, Daiichi silver, SYPRO Orange, SYPRO Red, SYPRO Ruby, or SYPRO Tangerine are compared. It has been concluded that the newest generation of fluorescent protein stains, compared with traditional staining methods, are more compatible to matrix-assisted laser desorption/ionization (MALDI) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods. The number of database matches obtained using each mass spectrometry method and the percent sequence coverage obtained from trypsin digested proteins stained using these six methods is provided.
Lauber
W M
WM
Pharmacia Corp, St Louis, MO 63198, USA. wendy.m.lauber@pharmacia.com
Carroll
J A
JA
Dufield
D R
DR
Kiesel
J R
JR
Radabaugh
M R
MR
Malone
J P
JP
D000595:Amino Acid Sequence; D000818:Animals; D001798:Blood Proteins / Q000032:analysis; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004396:Coloring Agents; D015180:Electrophoresis, Gel, Two-Dimensional; D005456:Fluorescent Dyes; D006801:Humans; D008099:Liver / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods*; D011506:Proteins / Q000032:analysis*; D051381:Rats; D012394:Rosaniline Dyes; D020539:Sequence Analysis, Protein; D016622:Silver Staining; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013194:Staining and Labeling / Q000379:methods; D000515:alpha 1-Antitrypsin / Q000032:analysis
D001798:Blood Proteins; D004396:Coloring Agents; D005456:Fluorescent Dyes; D011506:Proteins; D012394:Rosaniline Dyes; D000515:alpha 1-Antitrypsin; C048139:Coomassie blue