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Rapid and sensitive location of protein zones in gels using o‐phthaldialdehyde
Author(s) -
Taylor Michael D.,
Andrews Anthony T.
Publication year - 1981
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.1150020203
Subject(s) - reagent , chromatography , coomassie brilliant blue , agarose , chemistry , staining , glycoprotein , electrophoresis , fluorescence , polyacrylamide , size exclusion chromatography , enzyme , biochemistry , biology , genetics , physics , quantum mechanics , polymer chemistry
A new fluorescent staining technique using o‐phthaldialdehyde(OPA) has been developed for the rapid post‐electrophoretic location for separated zones of proteins and glycoproteins. The method has also been applied to the detection of immunoprecipitates and, in the form of a spray reagent, to the detection of proteins separated on thin‐layer gel filtration plates. Major protein zones can be visualized within 1–2 min with maximum sensitivity being achieved within 15 min. In polyacrylamide and agarose gels, the sensitivity is of the same order as staining with Coomassie Blue G‐250. Reagents are very inexpensive and, because the reaction is performed at neutral or weekly alkaline pH in aqueous buffers, retention of enzymatic or other biological activity should be optimal. The method is thus particularly suitable for use in preparative applications and when it is desirable to combine general protein detection with specific location of zone of enzyme activity.