{"id":1318,"date":"2020-12-03T14:19:37","date_gmt":"2020-12-03T05:19:37","guid":{"rendered":"https:\/\/www.promega.co.jp\/resources\/faq\/?p=1318"},"modified":"2021-01-19T14:50:32","modified_gmt":"2021-01-19T05:50:32","slug":"q592","status":"publish","type":"post","link":"https:\/\/www.promega.co.jp\/resources\/faq\/reporter-signaling\/cell-signaling\/kinase-assay\/kinase-glo\/q592","title":{"rendered":"Kinase-Glo\u00ae \u3067\u6e2c\u5b9a\u3067\u304d\u308b\u30ad\u30ca\u30fc\u30bc\u306b\u306f\u3069\u306e\u3088\u3046\u306a\u3082\u306e\u304c\u3042\u308a\u307e\u3059\u304b\uff1f"},"content":{"rendered":"\n<p>Kinase-Glo<sup>\u00ae<\/sup> \u306f\u30ad\u30ca\u30fc\u30bc\u53cd\u5fdc\u5f8c\u306e\u53cd\u5fdc\u6eb6\u6db2\u4e2d\u306eATP\u91cf\u3092\u6e2c\u5b9a\u3057\u307e\u3059\u306e\u3067\u3001\u3069\u306e\u30bf\u30a4\u30d7\u306e\u30ad\u30ca\u30fc\u30bc\u306b\u3082\u4f7f\u7528\u3067\u304d\u307e\u3059\u3002\u5f0a\u793e\u3067\u5b9f\u65bd\u3057\u305f\u30ad\u30ca\u30fc\u30bc\u306e\u53cd\u5fdc\u6761\u4ef6\u3092\u88681\u306b\u793a\u3057\u307e\u3059\u3002<\/p>\n\n\n\n<p>\u88681\u3000\u30ad\u30ca\u30fc\u30bc\u306e\u53cd\u5fdc\u6761\u4ef6<\/p>\n\n\n\n<figure class=\"wp-block-table\"><table><tbody><tr><th>Kinase<\/th><th>Substrate<\/th><th>ATP<\/th><th>Reaction Buffer<\/th><th>Kinase Rxn<br>Time\/Temp.<\/th><th>EC50<\/th><\/tr><tr><td>PKA<br>(V5161)<\/td><td>Kemptide (V5601) 5\u03bcM<\/td><td>1\u03bcM<\/td><td>40mM Tris [pH 7.5], 20mM MgCl<sub>2<\/sub>, 0.1mg\/mL BSA<\/td><td>20min., room temp.<\/td><td>0.15 units\/well<\/td><\/tr><tr><td>PKC<br>(V5261)<\/td><td>Neurogranin <sup>(28-43)<\/sup> (V5611) 10\u03bcM<\/td><td>1\u03bcM<\/td><td>20mM Tris [pH 7.5], 10mM MgCl<sub>2<\/sub>, 0.1mg\/mL BSA, 250uM EGTA, 400uM CaCl<sub>2<\/sub>, 0.32mg\/mL phosphatidylserine, 0.032mg\/mL diacylglycerol<\/td><td>90min., room temp.<\/td><td>7.9ng\/well<\/td><\/tr><tr><td>Lck<br>(Upstate, #14-442)<\/td><td>PTK peptide 2 substrate (V288A) 250\u03bcM<\/td><td>3\u03bcM<\/td><td>8mM imidazole hydrochloride [pH 7.3], 8mM beta-glycerophosphate, 200uM EGTA, 20mM MgCl<sub>2<\/sub>, 1mM MnCl<sub>2<\/sub>, 0.1mg\/mL BSA<\/td><td>60min., room temp.<\/td><td>4.3 m\u3000units\/well<\/td><\/tr><tr><td>Erk-2<\/td><td>66\u03bcM MBP (bovine myelin basic protein, Upstate, #13-104)<\/td><td>100nM<\/td><td>40mM Tris-HCl [pH 7.5], 0.1 mg\/mL BSA, 20mM MgCl<sub>2<\/sub><\/td><td>120min., room temp.<\/td><td>49ng\/well<\/td><\/tr><tr><td>PI 3-kinase (pp110Y)<\/td><td>0.5mg\/ml L-\u03b1-phosphatidylinositol (Sigma # P8443)<\/td><td>500nM<\/td><td>10mM Tris-HCl [pH 7.5], 50mM NaCl, 5mM MgCl<sub>2<\/sub><\/td><td>90min., room temp.<\/td><td>70ng\/well<\/td><\/tr><\/tbody><\/table><\/figure>\n","protected":false},"excerpt":{"rendered":"<p>Kinase-Glo\u00ae \u306f\u30ad\u30ca\u30fc\u30bc\u53cd\u5fdc\u5f8c\u306e\u53cd\u5fdc\u6eb6\u6db2\u4e2d\u306eATP\u91cf\u3092\u6e2c\u5b9a\u3057\u307e\u3059\u306e\u3067\u3001\u3069\u306e\u30bf\u30a4\u30d7\u306e\u30ad\u30ca\u30fc\u30bc\u306b\u3082\u4f7f\u7528\u3067\u304d\u307e\u3059\u3002\u5f0a\u793e\u3067\u5b9f\u65bd\u3057\u305f\u30ad\u30ca\u30fc\u30bc\u306e\u53cd\u5fdc\u6761\u4ef6\u3092\u88681\u306b\u793a\u3057\u307e\u3059\u3002 \u88681\u3000\u30ad\u30ca\u30fc\u30bc\u306e\u53cd\u5fdc\u6761\u4ef6 Kinase Subst &#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\/1318"}],"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=1318"}],"version-history":[{"count":1,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/posts\/1318\/revisions"}],"predecessor-version":[{"id":1319,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/posts\/1318\/revisions\/1319"}],"wp:attachment":[{"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/media?parent=1318"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/categories?post=1318"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/tags?post=1318"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}