{"id":1328,"date":"2020-12-03T14:22:15","date_gmt":"2020-12-03T05:22:15","guid":{"rendered":"https:\/\/www.promega.co.jp\/resources\/faq\/?p=1328"},"modified":"2021-01-19T14:56:54","modified_gmt":"2021-01-19T05:56:54","slug":"q597","status":"publish","type":"post","link":"https:\/\/www.promega.co.jp\/resources\/faq\/reporter-signaling\/cell-signaling\/kinase-assay\/kinase-glo\/q597","title":{"rendered":"\u57f9\u990a\u7d30\u80de\u62bd\u51fa\u6db2\u4e2d\u306e\u30ad\u30ca\u30fc\u30bc\u6d3b\u6027\u3092\u6e2c\u5b9a\u3067\u304d\u307e\u3059\u304b\uff1f"},"content":{"rendered":"\n<p>\u57f9\u990a\u7d30\u80de\u306b\u5b58\u5728\u3059\u308b\u30ad\u30ca\u30fc\u30bc\u306e\u6d3b\u6027\u3092Kinase-Glo<sup>\u00ae<\/sup> \u3067\u6e2c\u5b9a\u3059\u308b\u3053\u3068\u3082\u53ef\u80fd\u3067\u3059\u3002\u305f\u3060\u3057\u3001\u7d30\u80de\u306b\u306f\u591a\u7a2e\u985e\u306e\u30ad\u30ca\u30fc\u30bc\u304c\u542b\u307e\u308c\u308b\u305f\u3081\u4e00\u822c\u533b\u30d0\u30c3\u30af\u30b0\u30e9\u30a6\u30f3\u30c9\u304c\u9ad8\u304f\u3001\u307e\u305fATP\u5909\u5316\u304c\u6a19\u7684\u30ad\u30ca\u30fc\u30bc\u306b\u7531\u6765\u3059\u308b\u304b\u3069\u3046\u304b\u614e\u91cd\u306b\u691c\u8a0e\u3059\u308b\u5fc5\u8981\u304c\u3042\u308a\u307e\u3059\u3002<br>\u5b9f\u65bd\u4e8b\u4f8b\u3068\u3057\u3066U937\u7d30\u80de\u306b\u542b\u307e\u308c\u308bPKC(Protein Kinase C\uff09\u306e\u6d3b\u6027\u3092Kinase-Glo\u3092\u7528\u3044\u3066\u6e2c\u5b9a\u3057\u305f\u8ad6\u6587<sup>\uff083\uff09<\/sup>\u304c\u3042\u308a\u307e\u3059\u3002<br>\u6a19\u7684\u30ad\u30ca\u30fc\u30bc\u6d3b\u6027\u306e\u4f4e\u3044\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u306e\u7d30\u80de\u3068\u6a19\u7684\u30ad\u30ca\u30fc\u30bc\u6d3b\u6027\u306e\u9ad8\u3044\u30b5\u30f3\u30d7\u30eb\u306e\u7d30\u80de\u3092\u4f7f\u7528\u3057\u3066\u3001\u30ad\u30ca\u30fc\u30bc\u53cd\u5fdc\u5f8c\u306eATP\u91cf\u3092\u6bd4\u8f03\u3057\u3066\u3044\u307e\u3059\u3002<br>\u8ad6\u6587\u4e2d\u306e\u7d30\u80de\u62bd\u51fa\u65b9\u6cd5\u306f\u3001\u4ee5\u4e0b\u306e\u3068\u304a\u308a\u3067\u3059\u3002<\/p>\n\n\n\n<p>\u2460U937 cells \u30921 x 10<sup>7<\/sup>\u500b\u3068\u306a\u308b\u3088\u3046\u306b\u57f9\u990a\u3057\u307e\u3059\u3002<br>\u2461LPS\u30925\u03bcg\/mL\u3068\u306a\u308b\u3088\u3046\u306b\u57f9\u5730\u306b\u6dfb\u52a0\u3057\u300137\u2103\u30014 \u6642\u9593\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30c8\u3057\u307e\u3059\u3002<br>\u2462 \u51b7\u5374\u3057\u305fPBS\u3067\u7d30\u80de\u3092\u6d17\u6d44\u5f8c\u3001200 uL\u306ekinase reaction buffer (40 mM Tris, pH 7.5, 20 mM MgCl<sub>2<\/sub>, 0.1 mg\/mL BSA)\u3092\u6dfb\u52a0\u3057\u3001\u8d85\u97f3\u6ce2\u3067\u7d30\u80de\u3092\u7834\u7815\u3057\u307e\u3059\u3002<br>\u2463 \u30b5\u30f3\u30d7\u30eb\u309250uL\u306ePKC \u53cd\u5fdcbuffer (20 mM Tris, pH 7.5, 10 mM MgCl<sub>2<\/sub>, 0.1 mg\/mL BSA, 250 \u03bcM EGTA, 400 \u03bcM CaCl<sub>2<\/sub>, 0.32 mg\/ml phosphatidylserine, 0.032 mg\/mL diacylglycerol)\u3067\u5e0c\u91c8\u3057\u300110 \u03bcM \u306eATP \u3068100 \u03bcM\u306e neurogranin<sub>(28-43)<\/sub> \u3092\u6dfb\u52a0\u3057\u300190\u5206\u9593\u5ba4\u6e29\u3067\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30c8\u3057\u307e\u3059\u3002<br>\u246450\u03bcL\u306eKinase-Glo<sup>\u00ae<\/sup> \u8a66\u85ac\u3092\u6dfb\u52a0\u3057\u300110\u5206\u9593\u5ba4\u6e29\u3067\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30c8\u3057\u305f\u306e\u3061\u3001\u30eb\u30df\u30ce\u30e1\u30fc\u30bf\u30fc\u3067\u6e2c\u5b9a\u3057\u307e\u3059\u3002<\/p>\n\n\n\n<blockquote class=\"wp-block-quote\"><p>\u53c2\u8003\u6587\u732e(3):<br><em>Roman\u3000J. et al., Am J Physiol Lung Cell Mol Physiol 287: 287-249(2004)<\/em><\/p><\/blockquote>\n","protected":false},"excerpt":{"rendered":"<p>\u57f9\u990a\u7d30\u80de\u306b\u5b58\u5728\u3059\u308b\u30ad\u30ca\u30fc\u30bc\u306e\u6d3b\u6027\u3092Kinase-Glo\u00ae \u3067\u6e2c\u5b9a\u3059\u308b\u3053\u3068\u3082\u53ef\u80fd\u3067\u3059\u3002\u305f\u3060\u3057\u3001\u7d30\u80de\u306b\u306f\u591a\u7a2e\u985e\u306e\u30ad\u30ca\u30fc\u30bc\u304c\u542b\u307e\u308c\u308b\u305f\u3081\u4e00\u822c\u533b\u30d0\u30c3\u30af\u30b0\u30e9\u30a6\u30f3\u30c9\u304c\u9ad8\u304f\u3001\u307e\u305fATP\u5909\u5316\u304c\u6a19\u7684\u30ad\u30ca\u30fc\u30bc\u306b\u7531\u6765\u3059\u308b\u304b\u3069\u3046\u304b\u614e\u91cd\u306b\u691c\u8a0e\u3059\u308b &#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\/1328"}],"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=1328"}],"version-history":[{"count":1,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/posts\/1328\/revisions"}],"predecessor-version":[{"id":1329,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/posts\/1328\/revisions\/1329"}],"wp:attachment":[{"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/media?parent=1328"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/categories?post=1328"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/tags?post=1328"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}