通过酶联免疫吸附测定法对小鼠酪氨酸酶进行定量分析。
Quantitative analysis of mouse tyrosinase by enzyme-linked immunosorbent assay.
作者信息
Ishiguro S, Yamamoto H, Yanai N, Takeuchi T
出版信息
J Biochem. 1986 Apr;99(4):1081-5. doi: 10.1093/oxfordjournals.jbchem.a135572.
A sensitive, specific, competitive enzyme-linked immunosorbent assay (ELISA) was developed for quantitative analysis of tyrosinase. Binding sites of anti-tyrosinase antibodies were competed for by purified tyrosinase adsorbed onto microtiter plates and a known (standard) or unknown (sample) amount of tyrosinase in solution. Adsorbed antibodies were detected by goat anti-rabbit IgG F(ab')2 labeled with peroxidase. A sensitivity range of 2.1 to 14 ng (30-200 fmol)/well was obtained. SDS was found to be the most suitable detergent for solubilizing the enzyme. Tyrosinase was extracted from B16 mouse melanoma and assayed by the ELISA. The tyrosinase content per mg melanoma protein was 505 +/- 106 (S.D.) ng. This assay is not only useful for measuring the content of normal tyrosinase in crude extracts but also is possibly applicable to detecting the unprocessed tyrosinases.
开发了一种灵敏、特异的竞争性酶联免疫吸附测定法（ELISA）用于酪氨酸酶的定量分析。包被于微量滴定板上的纯化酪氨酸酶以及溶液中已知（标准品）或未知（样品）量的酪氨酸酶会竞争抗酪氨酸酶抗体的结合位点。用辣根过氧化物酶标记的山羊抗兔IgG F(ab')2检测吸附的抗体。获得了2.1至14 ng（30 - 200 fmol）/孔的灵敏度范围。发现SDS是溶解该酶最合适的去污剂。从B16小鼠黑色素瘤中提取酪氨酸酶并用ELISA进行测定。每毫克黑色素瘤蛋白中酪氨酸酶含量为505±106（标准差）ng。该测定法不仅可用于测量粗提物中正常酪氨酸酶的含量，还可能适用于检测未加工的酪氨酸酶。
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