Use of thiopropyl sepharose for the synthesis of an adsorbent for the affinity chromatography of glutathione S-transferase.
Thiopropyl Sepharose 6B in the 2-thiopyridyl-activated form was used for the reversible immobilisation of reduced glutathione (GSH). The resulting affinity matrix was successfully tested as a sorbent for the partial purification of glutathione S-transferase (GST) from pig kidney. The specific elution of the enzyme was performed with 10 mM GSH in Tris-HCl buffer (pH 7.8), non-specific elution with 20 mM dithiotreitol (DTT) in the same buffer.
Glatz
Z
Z
Department of Biochemistry, Masaryk University, Brno, Czech Republic.
Psotová
J
J
Janiczek
O
O
Chroust
K
K
Jowet
T
T
D000818:Animals; D002021:Buffers; D002846:Chromatography, Affinity / Q000379:methods; D005978:Glutathione / Q000737:chemistry; D005982:Glutathione Transferase / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D007668:Kidney / Q000201:enzymology; D012685:Sepharose / Q000031:analogs & derivatives* / Q000737:chemistry; D013552:Swine
D002021:Buffers; C035872:thiopropyl-sepharose; D012685:Sepharose; D005982:Glutathione Transferase; D005978:Glutathione