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A new method for the fast, reliable and reproducible submicrogram quantitation of proteins separated by different electrophoretic techniques is presented. The method is based on a modified sensitive staining technique using Coomassie Brilliant Blue G-250 in colloidal solution combined with an optimized elution procedure of the bound dye in a 3% w/v solution of sodium dodecyl sulfate followed by photometric determination of dye concentration in the eluate. In addition a new method is provided for background correction, even suitable for gels showing strong background staining. The staining procedure allows the detection of 20 ng depending on the nature of the protein and the separation technique used. Quantitation is linear at least in the range from 50 ng to 10 micrograms and highly reproducible even under non-optimized conditions. The presented method can be applied to sodium dodecyl sulfate-electrophoresis, isoelectric focusing and two-dimensional electrophoresis.
U Neumann, and
H Khalaf, and
M Rimpler
January 1987,
Caries research,
U Neumann, and
H Khalaf, and
M Rimpler
January 1996,
Methods in molecular biology (Clifton, N.J.),
U Neumann, and
H Khalaf, and
M Rimpler
April 1978,
Biochimica et biophysica acta,
U Neumann, and
H Khalaf, and
M Rimpler
January 1947,
The Biochemical journal,
U Neumann, and
H Khalaf, and
M Rimpler
January 1947,
The Biochemical journal,
U Neumann, and
H Khalaf, and
M Rimpler
May 1996,
Electrophoresis,
U Neumann, and
H Khalaf, and
M Rimpler
February 1977,
Medical research engineering,
U Neumann, and
H Khalaf, and
M Rimpler
August 1978,
The Japanese journal of experimental medicine,
U Neumann, and
H Khalaf, and
M Rimpler
January 1980,
Hoppe-Seyler's Zeitschrift fur physiologische Chemie,
U Neumann, and
H Khalaf, and
M Rimpler
January 1970,
Zeitschrift fur klinische Chemie und klinische Biochemie,
