A sensitive tyrosinase method for human skin.
A sensitive competitive radioimmunoassay for quantitative analysis of tyrosinase in biological tissues has been developed. The binding sites of antityrosinase antibodies were competed for by 125I-labelled human tyrosinase and a known (standard) or unknown (sample) amount of tyrosinase in solution. A sensitivity range of about 5-500 pmol/l was obtained when a solubilized melanoma tyrosinase with a molecular weight of 66,000 D was used as a standard. The method was used on 0.5 cm2 skin samples from which tyrosinase was solubilized with Nonidet P-40. The method is useful for measuring the content of tyrosinase in crude samples where the enzyme activity may be affected by inhibitors or by unspecific oxidation.
Department of Pharmacology, University Hospital, Lund, Sweden.
A
A
Wittbjer
G
G
Odh
E
E
Rosengren
H
H
Rorsman
D000818:Animals; D000918:Antibody Specificity / Q000276:immunology; D001666:Binding Sites, Antibody / Q000276:immunology; D001667:Binding, Competitive / Q000276:immunology; D002850:Chromatography, Gel / Q000379:methods; D006801:Humans; D066298:In Vitro Techniques; D007457:Iodine Radioisotopes; D008544:Melanocytes / Q000737:chemistry / Q000201:enzymology*; D014442:Monophenol Monooxygenase / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D011817:Rabbits; D011863:Radioimmunoassay / Q000379:methods*; D012867:Skin / Q000737:chemistry / Q000166:cytology / Q000201:enzymology*
D007457:Iodine Radioisotopes; D014442:Monophenol Monooxygenase