{"id":1324,"date":"2020-12-03T14:20:57","date_gmt":"2020-12-03T05:20:57","guid":{"rendered":"https:\/\/www.promega.co.jp\/resources\/faq\/?p=1324"},"modified":"2021-01-19T14:52:32","modified_gmt":"2021-01-19T05:52:32","slug":"q595","status":"publish","type":"post","link":"https:\/\/www.promega.co.jp\/resources\/faq\/reporter-signaling\/cell-signaling\/kinase-assay\/kinase-glo\/q595","title":{"rendered":"IC50\u306e\u7b97\u51fa\u65b9\u6cd5\u3092\u6559\u3048\u3066\u304f\u3060\u3055\u3044\u3002"},"content":{"rendered":"\n<p>IC50\u5024\u306e\u6e2c\u5b9a\u65b9\u6cd5\u3068\u3057\u3066\u3001\u30d7\u30ed\u30e1\u30ac\u3067\u306e\u5b9f\u65bd\u4f8b\u3092\u4ee5\u4e0b\u306b\u7d39\u4ecb\u3057\u307e\u3059\u3002<br>PKA (Cat.# V5161) \u30685\u03bcM Kemptide substrate (Cat.# V5601) \u309250\u03bcL \u306ekinase reaction buffer (40mM Tris [pH 7.5],20mM MgCl<sub>2<\/sub>, 0.1mg\/mL BSA) \u3067\u5e0c\u91c8\u3057\u3001ATP\u6fc3\u5ea6\u30921\u03bcM\u306b\u56fa\u5b9a\u3057\u307e\u3059\u3002\u30ad\u30ca\u30fc\u30bc\u53cd\u5fdc\u306f\u300120\u5206\u9593\u3001\u5ba4\u6e29\u3067\u5b9f\u65bd\u3057\u307e\u3057\u305f\u3002\u3055\u3089\u306b\u3001\u30ad\u30ca\u30fc\u30bc\u53cd\u5fdc\u6db2\u4e2d\u306b\u963b\u5bb3\u5264 (\uff28-8\u307e\u305f\u306fH-9) \u3092\u6dfb\u52a0\u3057\u307e\u3057\u305f\u3002<\/p>\n\n\n\n<p>\u963b\u5bb3\u5264\uff08H-8\u307e\u305f\u306f\u3001H-9\uff09\u306e\u6fc3\u5ea6\u3092\u5909\u5316\u3055\u305b\u305f\u3068\u304d\u306eRLU\u5024\u306e\u30d7\u30ed\u30c3\u30c8\u3092\u56f34\u306b\u793a\u3057\u307e\u3059\u3002IC<sub>50<\/sub>\u5024\u306f\u3001RLU\u5024\u306e\u4e2d\u9593\u5024\u306e\u5bfe\u6570\u5024\u6fc3\u5ea6\u304b\u3089\u7b97\u51fa\u3057\u307e\u3059\u3002\u66f2\u7dda\u8fd1\u4f3c\u306f\u3001GraphPad Prism<sup>\u00ae<\/sup>\u30b7\u30b0\u30e2\u30a4\u30c9\u7528\u91cf\u53cd\u5fdc\u66f2\u7dda(\u53ef\u5909\u52fe\u914d) \u7528\u30bd\u30d5\u30c8\u30a6\u30a7\u30a2\u3092\u7528\u3044\u3066\u884c\u3044\u307e\u3057\u305f\u3002<\/p>\n\n\n\n<p>\u56f34 A\u306f\u3001\u963b\u5bb3\u5264\u3068\u3057\u3066H-8\u3092\u4f7f\u7528\u3057\u305f\u5834\u5408\u306e\u30b0\u30e9\u30d5\u3067\u3059\u3002\u30d7\u30ed\u30c3\u30c8\u304b\u3089\u3001Log<sub>10<\/sub>IC<sub>50<\/sub>=0.56\u3088\u308a\u3001IC<sub>50<\/sub>=3.7\u03bcM\u3068\u306a\u308a\u307e\u3059\u3002\u56f34 B\u306f\u3001\u963b\u5bb3\u5264\u3068\u3057\u3066H-9\u3092\u4f7f\u7528\u3057\u305f\u3068\u304d\u306e\u3082\u306e\u3067\u3001Log<sub>10<\/sub>IC<sub>50<\/sub>=0.38\u3001IC<sub>50<\/sub>=2.4\u03bcM\u3068\u306a\u308a\u307e\u3059\u3002\u8ad6\u6587(1)\u3067\u5831\u544a\u3055\u308c\u3066\u3044\u308bIC<sub>50<\/sub>\u5024\u306f\u3001H-8\u3067\u306f\u3001IC<sub>50<\/sub>=1.2\u03bcM\u3001H-9\u3067\u306fIC<sub>50<\/sub>=1.9\u03bcM\u3068\u306a\u308a\u3001Kinase-Glo<sup>\u00ae<\/sup> \u3092\u7528\u3044\u3066\u7b97\u51fa\u3057\u305f\u5024\u306f\u3053\u308c\u3089\u306b\u8fd1\u3044\u5024\u3092\u793a\u3057\u3066\u3044\u307e\u3059\u3002<\/p>\n\n\n\n<figure class=\"wp-block-image size-large\"><img loading=\"lazy\" width=\"580\" height=\"240\" src=\"https:\/\/www.promega.co.jp\/resources\/faq\/wp-content\/uploads\/dia_kinase-glo_03.gif\" alt=\"\" class=\"wp-image-2264\"\/><figcaption>\u56f34 2\u7a2e\u985e\u306ePKA\u963b\u5bb3\u5264\u306e\u6ef4\u5b9a\u66f2\u7dda\u304b\u3089\u6c42\u3081\u308bIC<sub>50<\/sub>\u5024\u306e\u7b97\u51fa\u65b9\u6cd5<\/figcaption><\/figure>\n\n\n\n<blockquote class=\"wp-block-quote\"><p>IC50\u306e\u6c7a\u5b9a\u306b\u3064\u3044\u3066\u306f<a href=\"http:\/\/www.promega.com\/resources\/protocols\/technical-bulletins\/101\/kinase-glo-luminescent-kinase-assay-platform-protocol\/\">\u88fd\u54c1\u30d7\u30ed\u30c8\u30b3\u30eb<\/a>\u306e &#8220;Determining IC50 Values of Kinase Inhibitors&#8221; \u306e\u9805\u76ee\u3082\u3042\u308f\u305b\u3066\u3054\u53c2\u7167\u304f\u3060\u3055\u3044\u3002<\/p><p>\u53c2\u8003\u6587\u732e(1):<br><em>Hidaka, H., Watanabe, M. and Kobayashi, K. (1991) Properties and use of Hseriescompounds as protein kinase inhibitors. Methods in Enzymology, 201,328-339.<\/em><\/p><\/blockquote>\n","protected":false},"excerpt":{"rendered":"<p>IC50\u5024\u306e\u6e2c\u5b9a\u65b9\u6cd5\u3068\u3057\u3066\u3001\u30d7\u30ed\u30e1\u30ac\u3067\u306e\u5b9f\u65bd\u4f8b\u3092\u4ee5\u4e0b\u306b\u7d39\u4ecb\u3057\u307e\u3059\u3002PKA (Cat.# V5161) \u30685\u03bcM Kemptide substrate (Cat.# V5601) \u309250\u03bcL \u306ekinase reactio &#8230; <\/p>\n","protected":false},"author":16,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":[],"categories":[208],"tags":[],"_links":{"self":[{"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/posts\/1324"}],"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=1324"}],"version-history":[{"count":2,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/posts\/1324\/revisions"}],"predecessor-version":[{"id":2265,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/posts\/1324\/revisions\/2265"}],"wp:attachment":[{"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/media?parent=1324"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/categories?post=1324"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/tags?post=1324"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}