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Improved separation of alpha chains of collagen type I, type III, and type V by noninterrupted electrophoresis using thioglycolic acid as a negatively charged reducer
Author(s) -
Nakamura Kazuyuki,
Inoue ShinIchi,
Abiko Seishou,
Aoki Hideo,
Takeo Kazusuke
Publication year - 1989
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.1150100108
Subject(s) - thioglycolic acid , chemistry , electrophoresis , gel electrophoresis , chromatography , sodium dodecyl sulfate , monomer , polyacrylamide gel electrophoresis , buffer (optical fiber) , buffer solution , organic chemistry , biochemistry , polymer , enzyme , telecommunications , computer science
Improved separation of alpha chains of collagen type I (α 1 [I] 2 α 2 [I]), type III(α 1 [III] 3 ), and type V(α 1 [V] α 2 [V]α 3 [V]) was achieved by noninterrupted sodium dodecyl sulfate‐polyacrylamide gel electrophoresis with a negatively charged reducer, thioglycolic acid. The thioglycolic acid, added to the running buffer of the cathodic reservoir, in the middle of electrophoresis quickly migrated in the gel anode, reducing interchain disulfide linkages in collagen type III and dissociating it into its alpha chain monomer, α 1 [III], without an interruption of electrophoresis. The alpha chain, α 1 [III], migrated more slowly than the α 1 [I] and α 2 [I] chains of collagen type I, resulting in an excellent separation of α 1 [III] from α 1 [I]. The mobility of α 1 [III] could be controlled by varying the time of thioglycolic acid addition to the running buffer. This enabled us not only to separate α 1 [III] from α 1 [I] and α 1 [V], but also to precisely quantitate these alpha chains, even at low protein loading of mixed samples.