Electrophoretic separation and immunological identification of type 2X myosin heavy chain in rat skeletal muscle

doi:10.1016/0304-4165(90)90181-U

Biochimica et Biophysica Acta (BBA) - General Subjects

Volume 1035, Issue 1, 20 July 1990, Pages 109-112

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Abstract

One slow and three fast myosin heavry chains have been described in typical skeletal muscles of the adult rat using immunocytochemical analysis [1,2]. Electrophoretic isolation and immunochemical identification of these four isoforms has not been achieved. An electrophoretic procedure is described which, by altering the cross-linkage and polymerization kinetics of 5% polyacrylamide gels, allows resolution of these four distinct myosin heavy chains. Using specific monoclonal antibodies and double immunoblotting analysis, the identity and electrophoretic migration order of the myosin heavy chains was established to be: 2A < 2X < 2B < β/slow.

References (18)

P.J. Reiser et al.
J. Biol. chem.
(1985)
U. Carraro et al.
Biochem. Biophys. Res. Commun.
(1983)
A. Bar et al.
FEBS Lett.
(1988)
G.S. Butler-Browne et al.
Dev. Biol.
(1984)
S. Schiaffino et al.
J. Muscle Res. Cell Motil.
(1985)
S. Schiaffino et al.
S. Schiaffino et al.
Acta Physiol. Scand.
(1988)
J.I. Rushbrook et al.
D. Danieli-Betto et al.
Biochem. Biophys. Res. Commun.
(1986)

There are more references available in the full text version of this article.

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BBA reportElectrophoretic separation and immunological identification of type 2X myosin heavy chain in rat skeletal muscle

Abstract

J. Biol. chem.

Biochem. Biophys. Res. Commun.

FEBS Lett.

Dev. Biol.

J. Muscle Res. Cell Motil.

Acta Physiol. Scand.

Biochem. Biophys. Res. Commun.

BBA report
Electrophoretic separation and immunological identification of type 2X myosin heavy chain in rat skeletal muscle