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A rapid electrophoretic method for separating rabbit skeletal muscle myosin heavy chains at high resolution
Author(s) -
Kubis HansPeter,
Gros Gerolf
Publication year - 1997
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.1150180113
Subject(s) - myosin , electrophoresis , gel electrophoresis , major histocompatibility complex , chemistry , polyacrylamide gel electrophoresis , gene isoform , chromatography , skeletal muscle , microbiology and biotechnology , stacking , biochemistry , biology , anatomy , enzyme , gene , organic chemistry
An electrophoretic method using sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) was developed which gives a high‐resolution separation of the known myosin heavy chains of rabbit skeletal muscle with excellent reproducibility. The gel of 10 cm total length consists of (i) a first stacking gel of 3.5% total gel concentration (T) and pH 6.8, (ii) a first separating gel of 6.6%T and pH 8.8, (iii) a second stacking gel of 6.6%T and pH 6.8, and (iv) a second separating gel of 8.8%T and pH 8.8. With this composition, a minigel system allows separation of six myosin heavy‐chain (MHC) isoforms at room temperature without cooling and within 8h. In agreement with previous reports, the isoforms appear in the sequence MHC emb , MHC IIa, MHC IId, MHC neo , MHC IIb, MHC I. A special advantage is the detectability not only of the adult but also of the embryonic and neonatal isoforms MHC emb and MHC neo .