Isoelectric focusing of urinary metallothionein.
Isoelectric focusing of human urinary metallothionein at a pH range of 4.8 to 7.0 yielded a single protein band with a pI of 5.57 which co-migrated with authentic purified metallothionein I from human liver. Minimum pretreatment of the urine samples (160 ml) was needed. The preparatory steps included sample concentration with the original protein, enriched from 69 +/- 23 micrograms/ml to 2.0 +/- 1.4 mg/ml (+/- SD; n = 9), followed by heat treatment at 80 degrees C for 5 min (2.4 +/- 1.7 mg protein/ml). After focusing, the gels were stained with silver and the lanes were scanned with a laser scanner. Peak areas were used for quantitation with commercial beta 2-microglobulin as a standard. The urinary metallothionein ranged from 1.0 to 2.6 nmol/mmol creatinine, which is comparable with values reached by radio-immunoassay.
Institute of Occupational Health Sciences, University of Lausanne, Switzerland.
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Felley-Bosco
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PE
Hunziker
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H
Savolainen
D006358:Hot Temperature; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D007526:Isoelectric Point; D008668:Metallothionein / Q000652:urine*; D001613:beta 2-Microglobulin / Q000652:urine
D001613:beta 2-Microglobulin; D008668:Metallothionein