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Proteomics Workshop

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MCP Publication
Guidlines Draft

Overview of MCP
Guidelines for Clinical
Proteomics Manuscripts

MCP Guidelines
for Clinical Proteomics
Manuscripts

Contact Information:

MSCLS-St. Paul (map)
University of Minnesota
43 Gortner Laboratory
1479 Gortner Ave.
St. Paul, MN 55108
Phone: (612) 625-2280
Fax: (612) 625-5780

46 Gortner Laboratory
Phone: (612) 625-6283

MSCLS-Minneapolis (map)
University of Minnesota
5-194 MCB
420 Washington Ave.
Minneapolis, MN 55455
Phone: (612) 625-2279
or (612) 624-2674
Fax: (612) 625-2163

Email: webmaster

Destaining Protocols

Coomassie (CBB-R 250) Destaining Protocol

1. Cut band from the gel
2. Wash several times with 10 mM dithiotreitol (DTT), 0.1 M 4-ethylmorpholine acetate (pH 8.1) in 50% acetonitrile (ACN); (3 minutes in microwave oven per wash is suggested). Repeat until the Coomassie is completely removed.
3. Wash gel piece with water.
4. Shrink with 100% ACN (5 min sonicate)
5. Reswell with water (5 min sonicate)
6. Shrink with 100% ACN (5 min sonicate)
7. Reswell with water (5 min sonicate)
8. Wash with ACN:H₂O = 1:1 (5 min sonicate) (steps 3 - 7: salt and other contaminant removal)
9. Speedvac to near dryness.
10. Reconstitute sample with proteolytic digestion buffer→proteolytic digest

Method developed by Petr Novak at the Laboratory of Molecular Structure Characterization, Institute of Microbiology, Prague

Published in:

Karel Drbal, Pavla Angelisova, Ivan Hilgert, Jan Cerny, Petr Novak, and Vaclav Horejsi(2001) A proteolytically truncated form of free CD18, the common chain of leukocyte integrins, as a novel marker of activated myeloid cells. Blood, 98(5): 1561-6.

SYPRO (Bio-Rad Laboratories/Molecular Probes, Inc.) Ruby Protein Gel Stain Removal

1. After staining with Ruby Gel Stain excise protein band/spot from gel
2. Wash the gel slices 2 x 25 mM ammonium bicarbonate in 50% acetonitrile and 1 x 100% acetonitrile
3. Dry slices using a Speed Vac
4. Reconstitute sample with proteolytic digestion bufferà proteolytic digest

SYPRO destaining notes provided by Bio-Rad Laboratories Technical Services department (Aug 2001)