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A rapid detection system for the quantitative evaluation of radioactively labeled proteins in dehydrated slab gels
Author(s) -
Kruppa Joachim
Publication year - 1983
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150040504
Subject(s) - spectrum analyzer , polyacrylamide , chemistry , chromatography , polyacrylamide gel electrophoresis , linear relationship , biochemistry , optics , enzyme , physics , statistics , mathematics , polymer chemistry
3 H‐ and 35 S‐Labeled proteins of vesicular stomatitis virus were separated by polyacrylamide gel electrophoresis, localized and directly quantified by the Linear Analyzer after dehydration of the gel. The counting efficiency of the Linear Analyzer was approximately 0.1% for tritium in polyacrylamide gels, and increased after blotting to 2‐3%, whereas the detection efficiency for 35 S was 2‐3% in polyacrylamide gels. An increase in gel thickness reduced the detection efficiency of the Linear Analyzer significantly, an effect which was also observed in fluorography. The instrument can be used for direct, quantitative evaluation of tritium in gels and blots because a linear relationship exists between the input radioactivity and the recorded counts. Double‐label experiments with 3 H‐ and 35 S‐labeled proteins can also be quantified. The spatial resolution of the Linear Analyzer reaches the quality attained in fluorography. Quantitative evaluation of gels and blots by the Linear Analyzer has the additional advantage of being less time‐consuming than detection by fluorography.