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Standardization of protein position in silver‐stained two‐dimensional polyacrylamide gel electrophoresis
Author(s) -
Johnston Dennis A.,
Capetillo Sylvia,
Ramagli Louis S.,
Guevara Juan,
Gersten Douglas M.,
Rodriguez Lewis V.
Publication year - 1984
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.1150050209
Subject(s) - polyacrylamide gel electrophoresis , chromatography , gel electrophoresis of proteins , gel electrophoresis , two dimensional gel electrophoresis , electrophoresis , silver stain , standardization , polyacrylamide , chemistry , microbiology and biotechnology , biochemistry , biology , proteomics , computer science , enzyme , gene , operating system
Methods for standardization of silver‐“stained” protein position in two‐dimensional polyacrylamide gel electrophoresis are presented for their application to simultaneous multiple gel systems. Four methods are discussed: the use of a single gel per simultaneous run for standardization with either (1) known quantities of commercially prepared high‐or low‐molecular‐weight standards or (2) a known biological preparation such as T4 phage coat proteins; the use of each gel per simultaneous run to standardize itself by (3) including an internal carbamylation train such as creatine phosphokinase and using a one‐dimensional gel pattern of proteins such as rat heart whole homogenate along the margin(s) or (4) identifying “marker” protein spots in test samples that occur in all gels in a given study and standardizing to relative position. Method four is discussed in detail and examples given of its use in a comparative study of proteins in spontaneous primary hepatocellular carcinomas occurring in mice, and its use compared to the other methods.