SOLUBILIZATION OF PROTEIN-DYE COMPLEXES ON NITROCELLULOSE TO QUANTIFYPROTEINS SPECTROPHOTOMETRICALLY

Citation
Jpd. Goldring et L. Ravaioli, SOLUBILIZATION OF PROTEIN-DYE COMPLEXES ON NITROCELLULOSE TO QUANTIFYPROTEINS SPECTROPHOTOMETRICALLY, Analytical biochemistry, 242(2), 1996, pp. 197-201
Citations number
32
Categorie Soggetti
Biology
Journal title
ISSN journal
00032697
Volume
242
Issue
2
Year of publication
1996
Pages
197 - 201
Database
ISI
SICI code
0003-2697(1996)242:2<197:SOPCON>2.0.ZU;2-W
Abstract
Proteins absorbed directly onto nitrocellulose membranes were stained with amido black, ponceau S, colloidal silver, or Coomassie blue and s olubilized in dimethyl sulfoxide and the absorbance was measured spect rophotometrically. The optimal wavelength of each dye/protein/nitrocel lulose solution was found to be at 625, 529, 420, and 600 nm, respecti vely. A linear relationship was found between the protein concentratio n and absorbance at the appropriate wavelength for all the stains with individual purified proteins or protein mixtures. Protein (0.2-0.8 mu g) can be determined with the colloidal silver and 2-30 mu g with the other stains. Coomassie blue produced variable background staining of the nitrocellulose and is therefore not recommended. Proteins transfe rred electrophoretically to nitrocellulose from a sodium dodecyl sulfa te-polyacrylamide gel were also stained with the above dyes and solubi lized in dimethyl sulfoxide. Amido black was the most sensitive stain, detecting proteins in the range of 1-10 mu g. Components of the gel i nterfered with silver staining. (C) 1996 Academic Press, Inc.