{"id":998,"date":"2020-12-03T10:50:00","date_gmt":"2020-12-03T01:50:00","guid":{"rendered":"https:\/\/www.promega.co.jp\/resources\/faq\/?p=998"},"modified":"2021-01-27T16:53:21","modified_gmt":"2021-01-27T07:53:21","slug":"q748","status":"publish","type":"post","link":"https:\/\/www.promega.co.jp\/resources\/faq\/cell-health\/adme\/p450\/p450-glo\/q748","title":{"rendered":"P450-Glo\u2122\u3067\u6e2c\u5b9a\u3057\u305fRLU\u5024\u304b\u3089\u3001Km\u5024\u3084Ki\u5024\u3092\u6c42\u3081\u308b\u3053\u3068\u306f\u3067\u304d\u307e\u3059\u304b?"},"content":{"rendered":"\n

\u2460 Km\u5024
\u5f93\u6765\u306e\u65b9\u6cd5\u3069\u304a\u308a\u3001\u57fa\u8cea-\u53cd\u5fdc\u901f\u5ea6\u66f2\u7dda\u304b\u3089Km\u5024\u3092\u6c42\u3081\u308b\u3053\u3068\u304c\u3067\u304d\u307e\u3059\u3002Km\u5024\u3092\u6c42\u3081\u308b\u5834\u5408\u3001\u57fa\u8cea\u306b\u3088\u3063\u3066\u306f\u3001\u53cd\u5fdc\u901f\u5ea6\u304c\u98fd\u548c\u3059\u308b\u57fa\u8cea\u6fc3\u5ea6\u3088\u308a\u9ad8\u3044\u6fc3\u5ea6\u3067\u306f\u30eb\u30b7\u30d5\u30a7\u30e9\u30fc\u30bc\u306e\u53cd\u5fdc\u963b\u5bb3\u304c\u8d77\u3053\u308b\u3053\u3068\u304c\u78ba\u8a8d\u3055\u308c\u3066\u3044\u307e\u3059\uff08\u88682\uff09\u3002\u3088\u3063\u3066\u9ad8\u6fc3\u5ea6\u3067\u306e\u767a\u5149\u5024\u306f\u3001\u3055\u3089\u306b\u9ad8\u3044\u6fc3\u5ea6\u3067\u306e\u767a\u5149\u5024\u3068\u6bd4\u8f03\u3057\u3066\u53cd\u5fdc\u963b\u5bb3\u304c\u8d77\u3053\u3063\u3066\u3044\u306a\u3044\u3053\u3068\u3092\u78ba\u8a8d\u3059\u308b\u5fc5\u8981\u304c\u3042\u308a\u307e\u3059\u3002<\/p>\n\n\n\n

\u88682\u3000\u9ad8\u6fc3\u5ea6\u3067\u53cd\u5fdc\u901f\u5ea6\u306e\u98fd\u548c\u304c\u898b\u3089\u308c\u308bP450-Glo\u2122 Luciferin Substrate\u3068\u305d\u308c\u306b\u5bfe\u5fdc\u3059\u308bCYP450\u5206\u5b50\u7a2e<\/strong>
<\/p>\n\n\n\n

P450-Glo\u2122 Luciferin Substrate<\/strong><\/th>CYP450\u5206\u5b50\u7a2e<\/strong><\/th><\/tr>
Luciferin-ME<\/td>1A2, 2C8, 2C9, 2J2, 4A11, 4F3B, 19<\/td><\/tr>
Luciferin-CEE<\/td>1A1, 1B1, 3A7<\/td><\/tr>
Luciferin-H<\/td>2C9<\/td><\/tr>
Luciferin-ME EGE<\/td>2D6, 1A1, 1A2<\/td><\/tr>
Luciferin-H EGE<\/td>2C19, 1A1, 1A2<\/td><\/tr><\/tbody><\/table><\/figure>\n\n\n\n

\u30d7\u30ed\u30e1\u30ac\u3067\u6e2c\u5b9a\u3057\u305fP450-GloTM\u30a2\u30c3\u30bb\u30a4\u3067\u306eKm\u5024\u306f\u3001\u88fd\u54c1\u30de\u30cb\u30e5\u30a2\u30eb<\/a>\u306eTable2\u3001\u307e\u305f\u306f\u53c2\u8003\u6587\u732e(1)\u306b\u8a18\u8f09\u3055\u308c\u3066\u3044\u307e\u3059\u3002<\/p>\n\n\n\n

\u2461 Ki\u5024
Ki\u5024\u306e\u7b97\u51fa\u306f\u3001\u57fa\u8cea\u6fc3\u5ea6\u4e00\u5b9a\u3067\u3001\u963b\u5bb3\u5264\u306e\u6fc3\u5ea6\u3092\u5909\u66f4\u3057\u3066\u6e2c\u5b9a\u3059\u308b\u65b9\u6cd5\u3067\u7b97\u51fa\u3059\u308b\u3053\u3068\u304c\u3067\u304d\u307e\u3059\u3002
\u62ee\u6297\u963b\u5bb3\u5264\u306e\u5834\u5408\u3001Ki\u306f\u4ee5\u4e0b\u306e\u5f0f\u304b\u3089\u6c42\u3081\u308b\u3053\u3068\u304c\u3067\u304d\u307e\u3059\u3002<\/p>\n\n\n\n

Ki=IC50<\/sub>\/1+([substrate]\/Km)<\/p>\n\n\n\n

\u975e\u62ee\u6297\u963b\u5bb3\u5264\u306e\u5834\u5408\u3001Ki\u306f\u4ee5\u4e0b\u306e\u5f0f\u304b\u3089\u6c42\u3081\u308b\u3053\u3068\u304c\u3067\u304d\u307e\u3059\u3002<\/p>\n\n\n\n

Ki=[I]\/(Vmax\/Vimax)-1
[I]\uff1d\u963b\u5bb3\u5264\u6fc3\u5ea6\u3001Vimax\uff1d\u963b\u5bb3\u5264\u5b58\u5728\u4e0b\u3067\u306eVmax<\/p>\n\n\n\n

CYP2C9\u306e\u62ee\u6297\u963b\u5bb3\u5264sulfaphenazole\u306eKi\u5024\u3092\u6c42\u3081\u305f\u4f8b\u3092\u4ee5\u4e0b\u306b\u793a\u3057\u307e\u3059\u3002Luciferin-H\u306eKm=103\u03bcM\u3067\u3042\u308a(1)\u3001\u57fa\u8cea\u6fc3\u5ea6\u3092100\u03bcM\u3068\u3057\u305f\u3068\u304d\u963b\u5bb3\u5264\uff08sulfaphenazole\uff09\u306eIC50<\/sub>\u304c\u7d0410-7<\/sup>M=0.1\u03bcM\u3068\u306a\u308a\u307e\u3057\u305f\uff08\u56f32\u53c2\u7167\uff09\u3002\u3088\u3063\u3066Ki\u5024\u306f\u3001\u4ee5\u4e0b\u306e\u3088\u3046\u306b\u7b97\u51fa\u3055\u308c\u307e\u3059\u3002<\/p>\n\n\n\n

Ki=IC50<\/sub>\/1+([substrate]\/Km)
=0.1\/(1+{100\/103})=0.1\/1.97=0.05 (\u03bcM)<\/p>\n\n\n\n

\u56f32\u3000P450-Glo\u2122 Luciferin Substrate\uff08Luciferin-H\uff09\u6fc3\u5ea6\u3092\u4e00\u5b9a\u306b\u3057\u305f\u3068\u304d\u306e
sulfaphenazole\u306e\u6fc3\u5ea6\u5909\u5316\u306b\u3088\u308bCYP450\u6d3b\u6027\u963b\u5bb3\u306e\u5909\u5316\u91cf<\/strong><\/p>\n\n\n\n

\"\"<\/figure>\n\n\n\n

\u307e\u305f\u3001\u963b\u5bb3\u5264\u306e\u6fc3\u5ea6\u3092\u4e00\u5b9a\u306b\u3057\u3066\u3001\u57fa\u8cea\u6fc3\u5ea6\u3092\u5909\u66f4\u3059\u308b\u65b9\u6cd5\u3067\u3082Ki\u5024\u3092\u6c42\u3081\u308b\u3053\u3068\u304c\u3067\u304d\u307e\u3059\u3002<\/p>\n\n\n\n

\u53c2\u8003\u6587\u732e
1. Cali, J.S. et al. (2006) Luminogenic cytochrome P450 assays. Expert. Opin. Drug. Metab. Toxicol. 2, 629-45.<\/p>\n","protected":false},"excerpt":{"rendered":"

\u2460 Km\u5024\u5f93\u6765\u306e\u65b9\u6cd5\u3069\u304a\u308a\u3001\u57fa\u8cea-\u53cd\u5fdc\u901f\u5ea6\u66f2\u7dda\u304b\u3089Km\u5024\u3092\u6c42\u3081\u308b\u3053\u3068\u304c\u3067\u304d\u307e\u3059\u3002Km\u5024\u3092\u6c42\u3081\u308b\u5834\u5408\u3001\u57fa\u8cea\u306b\u3088\u3063\u3066\u306f\u3001\u53cd\u5fdc\u901f\u5ea6\u304c\u98fd\u548c\u3059\u308b\u57fa\u8cea\u6fc3\u5ea6\u3088\u308a\u9ad8\u3044\u6fc3\u5ea6\u3067\u306f\u30eb\u30b7\u30d5\u30a7\u30e9\u30fc\u30bc\u306e\u53cd\u5fdc\u963b\u5bb3\u304c\u8d77\u3053\u308b\u3053\u3068\u304c\u78ba\u8a8d\u3055\u308c\u3066\u3044\u307e\u3059\uff08\u88682\uff09 … <\/p>\n","protected":false},"author":16,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":[],"categories":[235],"tags":[],"_links":{"self":[{"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/posts\/998"}],"collection":[{"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/users\/16"}],"replies":[{"embeddable":true,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/comments?post=998"}],"version-history":[{"count":3,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/posts\/998\/revisions"}],"predecessor-version":[{"id":2462,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/posts\/998\/revisions\/2462"}],"wp:attachment":[{"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/media?parent=998"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/categories?post=998"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/tags?post=998"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}