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.
Wittbjer
A
A
Department of Pharmacology, University Hospital, Lund, Sweden.
Odh
G
G
Rosengren
E
E
Rorsman
H
H
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