Quantitation of Cu-containing metallothionein by a Cd-saturation method
- GSF Research Center, Neuherberg (Germany, F.R.)
A rapid and sensitive method for determining Cu-containing metallothionein (MT) is described. The main features of this Cd-saturation assay are: high-molecular-weight Cd-binding compounds are denatured with acetonitrile (50% final concentration), Cu bound to MT is removed with ammonium tetrathiomolybdate, excessive tetrathiomolybdate and its Cu complexes are removed with DEAE-Sephacel, apothionein is saturated with Cd, and excessive Cd is bound to Chelex 100. The thiomolybdate assay is capable of reliably detecting 14 ng MT and thus is particularly suitable for measuring MT in small tissue samples (e.g., biopsies), in extrahepatic tissues, and in cultured cells. Moreover, the combination of the thiomolybdate assay with the recently developed Cd-Chelex assay also makes it possible to determine the portion of MT which binds Cu (Cu load of MT), provided that the amount of non-Cu-thionein exceeds 100 ng, the detection limit of the Cd-Chelex assay.
- OSTI ID:
- 5889000
- Journal Information:
- Analytical Biochemistry; (USA), Vol. 189:1; ISSN 0003-2697
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
59 BASIC BIOLOGICAL SCIENCES
COPPER
METABOLISM
METALLOTHIONEIN
MEASURING METHODS
CADMIUM ISOTOPES
CHROMATOGRAPHY
FIBROBLASTS
GLUTATHIONE
MOLECULAR WEIGHT
RATS
TRACER TECHNIQUES
ANIMAL CELLS
ANIMALS
CONNECTIVE TISSUE CELLS
DRUGS
ELEMENTS
ISOTOPE APPLICATIONS
ISOTOPES
MAMMALS
METALLOPROTEINS
METALS
ORGANIC COMPOUNDS
PEPTIDES
POLYPEPTIDES
PROTEINS
RADIOPROTECTIVE SUBSTANCES
RODENTS
SEPARATION PROCESSES
SOMATIC CELLS
TRANSITION ELEMENTS
VERTEBRATES
560300* - Chemicals Metabolism & Toxicology
550501 - Metabolism- Tracer Techniques