Separation of metallothionein into isoforms by column switching on gel permeation and ion-exchange columns with high-performance liquid chromatography-atomic-absorption spectrophotometry.
Metallothionein in tissue supernatant was separated into isoforms by on-line switching from gel filtration to ion-exchange columns and the metals bound to the isoproteins were directly determined by using a flame atomic-absorption spectrophotometer as a specific detector for metals. Conditions for elution of the gel filtration column were established and Tris-HCl buffer solutions in low concentration were shown to be applicable to the column-switching experiment. The effects of pH, buffer and salt concentrations on the elution of metallothionein are discussed.
D000818:Animals; D002850:Chromatography, Gel / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D008099:Liver / Q000032:analysis; D008668:Metallothionein / Q000302:isolation & purification*; D051381:Rats; D011919:Rats, Inbred Strains; D013054:Spectrophotometry, Atomic / Q000379:methods; D013237:Stereoisomerism
D008668:Metallothionein