{"id":151,"date":"2015-08-03T13:56:00","date_gmt":"2015-08-03T04:56:00","guid":{"rendered":"http:\/\/www-p.cc.ocha.ac.jp\/bio-shimada-lab\/?page_id=151"},"modified":"2015-08-07T10:33:20","modified_gmt":"2015-08-07T01:33:20","slug":"pr_06protein","status":"publish","type":"page","link":"https:\/\/www-p.sci.ocha.ac.jp\/bio-shimada-lab\/ex_protocol\/pr_06protein.html","title":{"rendered":"\u30bf\u30f3\u30d1\u30af\u8cea\u95a2\u4fc2"},"content":{"rendered":"<h3>\u30bf\u30f3\u30d1\u30af\u8cea\u62bd\u51fa\u30d7\u30ed\u30c8\u30b3\u30eb<\/h3>\n<h4 style=\"text-align: left;\">\u30fb\u30fb\u30fb\u6e96\u5099\u6bb5\u968e\u30fb\u30fb\u30fb<\/h4>\n<p style=\"text-align: left;\">\u85fb\u4f53\u306f\u6c34\u6c17\u3092\u53d6\u3063\u3066\uff0c\u30b3\u30cb\u30ab\u30eb\u30c1\u30e5\u30fc\u30d6\u306b\u3044\u308c\uff0c\u6db2\u4f53\u7a92\u7d20\u3067\u77ac\u9593\u51b7\u51cd\u3057\uff0c-80\u3067\u4fdd\u5b58<\/p>\n<p style=\"text-align: left;\">\u53ef\u6eb6\u53ef\u30d0\u30c3\u30d5\u30a1\u30fc\u3000Triton-X, Tween 20\u3092\u6e96\u5099<\/p>\n<p style=\"text-align: left;\">\u30002.2\uff05TritonX-100\u53ef\u6eb6\u5316buffer<\/p>\n<ul>\n<li style=\"text-align: left;\">1M Tris-Cl (pH 7.5) 250\u03bcl [50mM]<\/li>\n<li style=\"text-align: left;\">2-\u30e1\u30eb\u30ab\u30d7\u30c8\u30a8\u30bf\u30ce\u30fc\u30eb 110\u03bcl [0.31M]<\/li>\n<li style=\"text-align: left;\">10% TritonX-100 1.1ml [2.2%(v\/v)]<\/li>\n<li style=\"text-align: left;\">137mM PMSF 292\u03bcl [8mM]<\/li>\n<\/ul>\n<table style=\"height: 25px;\" width=\"272\">\n<tbody>\n<tr>\n<td>Fill up to 5ml (DW)<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p style=\"text-align: left;\">\n<h4 style=\"text-align: left;\">Tween-20 (Polyoxethylene(20)sorbitan monolaurate)\u53ef\u6eb6\u5316buffer<\/h4>\n<ul>\n<li style=\"text-align: left;\">1M Tris-Cl (pH 7.5) 250\u03bcl [50mM]<\/li>\n<li style=\"text-align: left;\">2-\u30e1\u30eb\u30ab\u30d7\u30c8\u30a8\u30bf\u30ce\u30fc\u30eb 110\u03bcl [0.31M]<\/li>\n<li style=\"text-align: left;\">5% Tween-20 1ml [1%(v\/v)]<\/li>\n<li style=\"text-align: left;\">137mM PMSF 292\u03bcl [8mM]<\/li>\n<\/ul>\n<table>\n<tbody>\n<tr>\n<td width=\"360\">Fill up to 5ml (DW)<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p style=\"text-align: left;\">\n<h4 style=\"text-align: left;\">L solution<\/h4>\n<ul>\n<li style=\"text-align: left;\">Tris 36.4g [1.5M]<\/li>\n<li style=\"text-align: left;\">\u30fb10%SDS 8ml [0.4M]<\/li>\n<\/ul>\n<table>\n<tbody>\n<tr>\n<td width=\"253\">Fill up to 200ml (DW)<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p style=\"text-align: left;\">\u3000\u4e0a\u8a18\u306e\u8a66\u85ac\u3092DW\u306b\u6eb6\u304b\u3057200ml\u8fd1\u304f\u307e\u3067fill up\u3057\u3001conc.HCl\u3092\u7528\u3044\u3066pH8.8\u306b\u8abf\u6574\u3057\u305f\u3002<\/p>\n<h4 style=\"text-align: left;\">M solution<\/h4>\n<ul>\n<li style=\"text-align: left;\">Tris\u3000\u3000\u3000\u3000 10g [0.5M]<\/li>\n<li style=\"text-align: left;\">10%SDS 8ml [0.4M]<\/li>\n<\/ul>\n<table>\n<tbody>\n<tr>\n<td width=\"256\">Fill up to 200ml (DW)<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p style=\"text-align: left;\">\u4e0a\u8a18\u306e\u8a66\u85ac\u3092DW\u306b\u6eb6\u304b\u3057200ml\u8fd1\u304f\u307e\u3067fill up\u3057\u3001conc.HCl\u3092\u7528\u3044\u3066pH6.8\u306b\u8abf\u6574\u3057\u305f\u3002<\/p>\n<h4 style=\"text-align: left;\">N solution<\/h4>\n<ul>\n<li style=\"text-align: left;\">Acrylamide\u3000 60g [30%(w\/v)]<\/li>\n<li style=\"text-align: left;\">N,N\u2019-Methylene-bis-acrylamide 2.16g [1.08%(w\/v)]<\/li>\n<\/ul>\n<table>\n<tbody>\n<tr>\n<td width=\"412\">Fill up to 200ml (DW)<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<ul>\n<li style=\"text-align: left;\">10% Ammonium peroxdisulfate (APS)<\/li>\n<li style=\"text-align: left;\">-20\u2103\u3067\u4fdd\u5b58\u3002<\/li>\n<li style=\"text-align: left;\">10\u00d7Running buffer<\/li>\n<li style=\"text-align: left;\">Tris\u3000\u3000\u3000\u3000\u3000\u3000 30g[250mM]<\/li>\n<li style=\"text-align: left;\">Glycine 144g[1.92M]<\/li>\n<\/ul>\n<table>\n<tbody>\n<tr>\n<td width=\"284\">SDS 10g[1%(w\/v)]<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p style=\"text-align: left;\">Fill up to 1 liter (DW)<\/p>\n<h4 style=\"text-align: left;\">\u2606\u30b2\u30eb\u88c5\u7f6e\u306e\u7d44\u307f\u7acb\u3066\u2606<\/h4>\n<ul>\n<li style=\"text-align: left;\">\u30b2\u30eb\u677f\uff0c\u30b3\u30fc\u30e0\uff0c\u30b4\u30e0\u30d0\u30f3\u30c9\u3092\u6d17\u6d44\uff08\u624b\u888b\u3092\u5fd8\u308c\u305a\u306b\uff09\u3002<\/li>\n<li>DW\u3092\u304b\u3051\uff0c\u30ad\u30e0\u30ef\u30a4\u30d7\u3067\u62ed\u304d\uff0c\u65b0\u3057\u3044\u30ad\u30e0\u30bf\u30aa\u30eb\u306e\u4e0a\u306b\u7f6e\u304f\u3002<\/li>\n<li>Gel module\u306b\u30ac\u30e9\u30b9\u30d7\u30ec\u30fc\u30c8\u3068\u30b9\u30da\u30b5\u30fc\u3092\u30bb\u30c3\u30c8\u3059\u308b\uff08\u51f9\u677f\u3092\u4e0b\u5074\u306b\u3059\u308b\uff09\u3002<\/li>\n<li>\u4e21\u30b5\u30a4\u30c9\u3092\u30af\u30e9\u30f3\u30d7\u3067\u631f\u307f\u3001\u30b9\u30af\u30ea\u30e5\u30fc\u3092\u5747\u7b49\u306b\u7de0\u3081\u308b\u3002\u30b7\u30fc\u30ea\u30f3\u30b0\u30d7\u30ec\u30fc\u30c8\u3092\u8d77\u3053\u3057\u3001\u3064\u307e\u307f\u3092\u30af\u30e9\u30f3\u30d7\u306e\u5965\u306e\u7aaa\u307f\u306b\u306f\u3081\u8fbc\u3080\u3002<\/li>\n<li>\u30b3\u30fc\u30e0\u3092\u3055\u3057\uff0c\u305d\u306e2cm\u4e0b\u306b\u77e2\u5370\u30b7\u30fc\u30eb\u3092\u306f\u308b\u3002<\/li>\n<li>70%\u30a8\u30bf\u30ce\u30fc\u30eb\u3092\u3044\u308c\u3066\u307f\u3066\u6f0f\u308c\u304c\u306a\u3044\u304b\u30c1\u30a7\u30c3\u30af\u3002<\/li>\n<\/ul>\n<p style=\"text-align: left;\">\u2606\u30b2\u30eb\u306e\u4f5c\u88fd\u2606<\/p>\n<ul>\n<li style=\"text-align: left;\">Running\u30b2\u30eb\u306e\u6750\u6599\u3092\u4e09\u89d2\u30d5\u30e9\u30b9\u30b3\u306b\u3044\u308c\uff0c\u624b\u3067\u56de\u3057\u307e\u305c\u308b\uff08\u4e0b\u8868\u53c2\u7167\uff09\u3002<\/li>\n<li>\u8131\u6c17\u306f\u305b\u305a\u306b\uff0c\u305d\u306e\u307e\u307e\u77e2\u5370\u30b7\u30fc\u30eb\u306e\u3061\u3087\u3044\u4e0a\u307e\u3067\u6d41\u3057\u8fbc\u3080\u3002<\/li>\n<li>70%\u30a8\u30bf\u30ce\u30fc\u30eb\u3092\u5165\u308c\uff0c\u30ab\u30d0\u30fc\u3059\u308b\u3002<\/li>\n<li>\u30e9\u30c3\u30d7\u3067\u4e0a\u90e8\u3092\u3064\u3064\u307f\uff0c37\u2103, 15min.<\/li>\n<li style=\"text-align: left;\">Stacking\u30b2\u30eb\u306e\u6750\u6599\u3092\u4e09\u89d2\u30d5\u30e9\u30b9\u30b3\u306b\u3044\u308c\uff0c\u624b\u3067\u56de\u3057\u307e\u305c\u308b\uff08\u4e0b\u8868\u53c2\u7167\uff09\u3002<\/li>\n<li>\u30b2\u30eb\u4e0a\u90e8\u306e70%\u30a8\u30bf\u30ce\u30fc\u30eb\u3092\u6368\u3066\uff0c\u308d\u7d19\u3067\u3057\u3063\u304b\u308a\u3068\u5438\u3044\u53d6\u308b\u3002<\/li>\n<li>Stacking\u30b2\u30eb\u3092\u304e\u308a\u304e\u308a\u307e\u3067\u3044\u308c\uff0c\u9759\u304b\u306b\u30b3\u30fc\u30e0\u3092\u5dee\u3059\uff08\u6ce1\u304c\u306a\u3044\u304b\u78ba\u8a8d\uff09\u3002<\/li>\n<li>\u30e9\u30c3\u30d7\u3067\u4e0a\u90e8\u3092\u3064\u3064\u307f\uff0c37\u2103, 20min.<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n<h4>7.5% Running gel<\/h4>\n<table>\n<tbody>\n<tr>\n<th><\/th>\n<th style=\"text-align: center;\">\u4e00\u679a\u5206<\/th>\n<th style=\"text-align: center;\">\u4e8c\u679a\u5206<\/th>\n<\/tr>\n<tr>\n<td style=\"text-align: left;\">DW<\/td>\n<td style=\"text-align: center;\">4.976ml<\/td>\n<td style=\"text-align: center;\">9.952ml<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: left;\">L solution<\/td>\n<td style=\"text-align: center;\">\u00a02.49ml<\/td>\n<td style=\"text-align: center;\">\u00a04.976ml<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: left;\">N solution<\/td>\n<td style=\"text-align: center;\">4.976ml<\/td>\n<td style=\"text-align: center;\"><\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: left;\">10% APS<\/td>\n<td style=\"text-align: center;\">39.8\u03bcl<\/td>\n<td style=\"text-align: center;\">79.6\u03bcl<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: left;\">TEMED<\/td>\n<td style=\"text-align: center;\">10\u03bcl<\/td>\n<td style=\"text-align: center;\">20\u03bcl<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<h4 style=\"text-align: left;\">10% Running gel<\/h4>\n<table>\n<tbody>\n<tr>\n<th style=\"text-align: left;\"><\/th>\n<th style=\"text-align: center;\">\u4e00\u679a\u5206<\/th>\n<th style=\"text-align: center;\">\u4e8c\u679a\u5206<\/th>\n<\/tr>\n<tr>\n<td style=\"text-align: left;\">DW<\/td>\n<td style=\"text-align: center;\">\u00a04.1ml<\/td>\n<td style=\"text-align: center;\">8.2ml<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: left;\">L solution<\/td>\n<td style=\"text-align: center;\">\u00a02.49ml<\/td>\n<td style=\"text-align: center;\">\u00a04.98ml<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: left;\">N solution<\/td>\n<td style=\"text-align: center;\">\u00a03.3ml<\/td>\n<td style=\"text-align: center;\">\u00a06.6ml<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: left;\">10% APS<\/td>\n<td style=\"text-align: center;\">39.8\u03bcl<\/td>\n<td style=\"text-align: center;\">79.6\u03bcl<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: left;\">TEMED<\/td>\n<td style=\"text-align: center;\">10\u03bcl<\/td>\n<td style=\"text-align: center;\">20\u03bcl<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p>&nbsp;<\/p>\n<h4>12.5% running gel<\/h4>\n<table>\n<tbody>\n<tr>\n<th><\/th>\n<th style=\"text-align: center;\">\u4e00\u679a\u5206<\/th>\n<th style=\"text-align: center;\">\u4e8c\u679a\u5206<\/th>\n<\/tr>\n<tr>\n<td>DW<\/td>\n<td style=\"text-align: center;\">3.39ml<\/td>\n<td style=\"text-align: center;\">6.78ml<\/td>\n<\/tr>\n<tr>\n<td>L solution<\/td>\n<td style=\"text-align: center;\">2.49ml<\/td>\n<td style=\"text-align: center;\">4.98ml<\/td>\n<\/tr>\n<tr>\n<td>N solution<\/td>\n<td style=\"text-align: center;\">4.17ml<\/td>\n<td style=\"text-align: center;\">8.34ml<\/td>\n<\/tr>\n<tr>\n<td>10% APS<\/td>\n<td style=\"text-align: center;\">39.8\u03bcl<\/td>\n<td style=\"text-align: center;\">79.6\u03bcl<\/td>\n<\/tr>\n<tr>\n<td>TEMED<\/td>\n<td style=\"text-align: center;\">10\u03bcl<\/td>\n<td style=\"text-align: center;\">20\u03bcl<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p>&nbsp;<\/p>\n<h4>15% running gel<\/h4>\n<table>\n<tbody>\n<tr>\n<th><\/th>\n<th style=\"text-align: center;\">\u4e00\u679a\u5206<\/th>\n<th style=\"text-align: center;\">\u4e8c\u679a\u5206<\/th>\n<\/tr>\n<tr>\n<td>DW<\/td>\n<td>2.488ml<\/td>\n<td>4.976ml<\/td>\n<\/tr>\n<tr>\n<td>L solution<\/td>\n<td>2.488ml<\/td>\n<td>4.976ml<\/td>\n<\/tr>\n<tr>\n<td>N solution<\/td>\n<td>4.976ml<\/td>\n<td>9.952ml<\/td>\n<\/tr>\n<tr>\n<td>10% APS<\/td>\n<td>39.8\u03bcl<\/td>\n<td>79.6\u03bcl<\/td>\n<\/tr>\n<tr>\n<td>TEMED<\/td>\n<td>10\u03bcl<\/td>\n<td>20\u03bcl<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<h4 style=\"text-align: left;\">stacking gel<\/h4>\n<table>\n<tbody>\n<tr>\n<th><\/th>\n<th style=\"text-align: center;\">\u4e00\u679a\u5206<\/th>\n<th style=\"text-align: center;\">\u4e8c\u679a\u5206<\/th>\n<\/tr>\n<tr>\n<td>DW<\/td>\n<td style=\"text-align: center;\">2.98ml<\/td>\n<td style=\"text-align: center;\">5.96ml<\/td>\n<\/tr>\n<tr>\n<td>M solution<\/td>\n<td style=\"text-align: center;\">1.26ml<\/td>\n<td style=\"text-align: center;\">2.52ml<\/td>\n<\/tr>\n<tr>\n<td>N solution<\/td>\n<td style=\"text-align: center;\">0.77ml<\/td>\n<td style=\"text-align: center;\">1.54ml<\/td>\n<\/tr>\n<tr>\n<td>10% APS<\/td>\n<td style=\"text-align: center;\">16.8\u03bcl<\/td>\n<td style=\"text-align: center;\">33.6\u03bcl<\/td>\n<\/tr>\n<tr>\n<td>TEMED<\/td>\n<td style=\"text-align: center;\">10\u03bcl<\/td>\n<td style=\"text-align: center;\">20\u03bcl<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<h4 style=\"text-align: left;\">\u2606\u30bf\u30f3\u30d1\u30af\u8cea\u306e\u62bd\u51fa\u2606<\/h4>\n<ul>\n<li>\u4e73\u9262\u306b0.2g\u306e\u85fb\u4f53\u3092\u3068\u308a\uff0c\u6c37\u4e0a\u3067\u53ef\u6eb6\u53ef\u30d0\u30c3\u30d5\u30a1\u30fc\u3092800\u03bcl\u5165\u308c\u7c89\u7815\u3059\u308b\u3002<\/li>\n<li>5ml\u30c1\u30e5\u30fc\u30d6\u306b\u5168\u91cf\u5165\u308c\uff0c3000rpm, 10min<\/li>\n<li>\u4e0a\u6f84\u307f\u3092\u5225\u306e\u30c1\u30e5\u30fc\u30d6\u306b\u5168\u91cf\u5165\u308c\uff0c\u6c37\u4e0a<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n<h4 style=\"text-align: left;\">\u2606\u30ed\u30fc\u30c7\u30a3\u30f3\u30b0\u2606<\/h4>\n<p style=\"text-align: left;\">4\u00d7sample buffer<\/p>\n<p style=\"text-align: left;\">0.5 M Tris-HCl pH 6.8 5ml [0.5M]<\/p>\n<p style=\"text-align: left;\">SDS 2.4g [12%(w\/v)]<\/p>\n<p style=\"text-align: left;\">Glycerol 8ml [40%(v\/v)]<\/p>\n<p style=\"text-align: left;\">2-mercapto ethanol 4ml [20%(v\/v)]<\/p>\n<table>\n<tbody>\n<tr>\n<td width=\"397\">Bromophenol blue 0.8mg [0.004%(w\/v)]<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p style=\"text-align: left;\">Fill up to 20ml (DW)<\/p>\n<ul>\n<li style=\"text-align: left;\">Gel module\u306e\u30b7\u30fc\u30ea\u30f3\u30b0\u30d7\u30ec\u30fc\u30c8\u3092\u958b\u3044\u3066\u3001\u91cd\u5408\u3057\u3066\u3044\u306a\u3044\u30b2\u30eb\u6eb6\u6db2\u3092\u308d\u7d19\u3067\u3075\u304d\u53d6\u308b\u3002<\/li>\n<li style=\"text-align: left;\">\u30b7\u30fc\u30ea\u30f3\u30b0\u30d7\u30ec\u30fc\u30c8\u306e\u3064\u307e\u307f\u3092\u30af\u30e9\u30f3\u30d7\u306e\u4e0b\u5074\u306e\u7aaa\u307f\u306b\u306f\u3081\u8fbc\u3080\u3002<\/li>\n<li style=\"text-align: left;\">\u62bd\u51fa\u6db215\u03bcl, 4x\u30d0\u30c3\u30d5\u30a1\u30fc5\u03bcl\uff08updpwn\uff09\u3092PCR\u30c1\u30e5\u30fc\u30d6\u306b\u3068\u308a\uff0cpipetting\u3002<\/li>\n<li style=\"text-align: left;\">100\u2103, 5min\u3067\u30d2\u30fc\u30c8\u30b7\u30e7\u30c3\u30af\uff0c\u30b9\u30d4\u30f3\u30c0\u30a6\u30f3\uff08\u673a\u4e0a\uff09\u3002<\/li>\n<li>\u30b3\u30fc\u30e0\u3092\u3086\u3063\u304f\u308a\u306f\u305a\u3057\uff0c\u7d30\u304f\u5207\u3063\u305f\u308d\u7d19\u3067cell\u3092\u304d\u308c\u3044\u306b\u3002<\/li>\n<li>Running\u30d0\u30c3\u30d5\u30a1\u30fc\u3092\u4f5c\u88fd\u3002<\/li>\n<li>\u5916\u5074\u306b\uff11\uff0f\uff13\u3050\u3089\u3044Running\u30d0\u30c3\u30d5\u30a1\u30fc\u3092\u3044\u308c\u308b\u3002<\/li>\n<li>Gel module\u3092\u88c5\u7740\uff08\u4e0b\u306b\u6ce1\u3092\u3044\u308c\u306a\u3044\u3088\u3046\u306b\uff09\u3002\u4e0b\u306b\u6ce1\u304c\u5165\u3063\u305f\u5834\u5408\u306b\u306f\u30b7\u30ea\u30f3\u30b8\u3092\u7528\u3044\u3066\u6ce1\u3092\u53d6\u308a\u9664\u304f\u3002<\/li>\n<li>\u5185\u5074\u306bRunning\u30d0\u30c3\u30d5\u30a1\u30fc\u3092\u3044\u308c\u308b\u3002<\/li>\n<li>16\u03bcl\u3092\u30ed\u30fc\u30c7\u30a3\u30f3\u30b0\u3002\u30de\u30fc\u30ab\u306f5\u03bcl<\/li>\n<li>1\u679a\u306710mA\uff0c\u88cf\u8868\u306720mA\u3001\u4e0b\u30b2\u30eb\u306b\u9054\u3057\u305f\u30891\u679a\u306720mA\uff0c\u88cf\u8868\u306740mA<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n<h4 style=\"text-align: left;\">\u2606\u67d3\u8272\u2606<\/h4>\n<ul>\n<li style=\"text-align: left;\">\u30d0\u30f3\u30c9\u304c\u4e0b\u307e\u3067\u9054\u3057sample buffer\u304crunning buffer\u306b\u51fa\u3060\u3057\u305f\u3068\u3053\u308d\u3067\uff0c\uff10\u306b\u623b\u3057\uff0c\u96fb\u6e90off\u3002<\/li>\n<li>\u30ac\u30e9\u30b9\u677f\u3092\u30b9\u30d1\u30c1\u30e5\u30e9\u3067\u306f\u305a\u3057\uff0cStacking\u30b2\u30eb\u3092\u5207\u308a\u53d6\u308b\u3002<\/li>\n<li>\u30bf\u30c3\u30d1\u306bDW\u3092\u5165\u308c\uff0c\u305d\u3053\u306bDW\u3092\u304b\u3051\u306a\u304c\u3089\u30b2\u30eb\u3092\u6d41\u3057\u5165\u308c10min\u3057\u3093\u3068\u3046\u3002<\/li>\n<li>DW\u3092\u3059\u3066\uff0c CBB\u67d3\u8272\u6db2\u3092\u3044\u308c\uff0c30min\u3057\u3093\u3068\u3046\u3002<\/li>\n<li>\u67d3\u8272\u6db2\u3092\u4fdd\u5b58\u7528\u30d3\u30f3\u306b\u623b\u3057\uff0c\u8131\u8272\u6db2\u3092\u9069\u91cf\u5165\u308c\uff0c\u30ad\u30e0\u30ef\u30a4\u30d7\u3092\u5165\u308c30min\u3057\u3093\u3068\u3046\u3002<\/li>\n<li>\u5ec3\u6db2\u3092\u5ec3\u6db2\u5165\u308c\u306b\u5165\u308c\uff0c\u30bf\u30c3\u30d1\u3092\u6d17\u6d44\u3002<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n<h4 style=\"text-align: left;\">\u67d3\u8272\u5e0c\u91c8\u30fb\u8131\u8272\u6db2<\/h4>\n<p style=\"text-align: left;\">\u7279\u7d1aMeOH 300ml [30%(v\/v)]<\/p>\n<table>\n<tbody>\n<tr>\n<td width=\"300\">\u7279\u7d1aAcetic Acid 100ml [10%(v\/v)]<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p style=\"text-align: left;\">Fill up to 1 liter (DW)<\/p>\n<h4 style=\"text-align: left;\">10\u00d7\u67d3\u8272\u6db2<\/h4>\n<p style=\"text-align: left;\">\u7279\u7d1aMeOH 30ml [30%(v\/v)]<\/p>\n<table>\n<tbody>\n<tr>\n<td width=\"300\">\u7279\u7d1aAcetic Acid 10ml [10%(v\/v)]<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p style=\"text-align: left;\">Fill up to 100 ml (DW)<\/p>\n<p style=\"text-align: left;\">\u2192200ml\u5bb9\u30d3\u30fc\u30ab\u30fc\u306b\u8abf\u6574\u3057250mg\u306eCoomassie Brilliant Blue R-250 (CBB)\u3092\u5165\u308c\u3066\u6eb6\u89e3\u3055\u305b\u308b\u3002<\/p>\n<h4 style=\"text-align: left;\">\u2606\u30b2\u30eb\u4e7e\u71e5\u2606<\/h4>\n<ul>\n<li style=\"text-align: left;\">\u30a2\u30c3\u30bb\u30f3\u30d6\u30ea\u30fc\u30c6\u30fc\u30d6\u30eb\u306b\u30dc\u30c8\u30e0\u30d5\u30ec\u30fc\u30e0\u3068\u6c34\u3067\u306c\u3089\u3057\u305f\u30bb\u30ed\u30d5\u30a1\u30f3\u3092\u306e\u305b\u308b\u3002<\/li>\n<li>\u30bb\u30ed\u30d5\u30a1\u30f3\u306e\u4e0a\u306b\u30b2\u30eb\u3092\u306e\u305b\u308b\u3002<\/li>\n<li>\u30b2\u30eb\u306e\u4e0a\u306b\u3082\u3046\u4e00\u679a\u6c34\u3067\u306c\u3089\u3057\u305f\u30bb\u30ed\u30d5\u30a1\u30f3\u3092\u306e\u305b\u308b\u3002\u3053\u306e\u3068\u304d\u7a7a\u6c17\u304c\u5165\u3089\u306a\u3044\u3088\u3046\u306b\u6ce8\u610f\u3059\u308b\u3053\u3068\u3002<\/li>\n<li>\u30c8\u30c3\u30d7\u30d5\u30ec\u30fc\u30e0\u3092\u306e\u305b\u3066\u30af\u30e9\u30f3\u30d7\u3067\u7de0\u3081\u308b\u3002<\/li>\n<li>\u30c9\u30e9\u30a4\u30e4\u30fc\u306e\u4e2d\u306b\u30d5\u30ec\u30fc\u30e0\u3092\u5165\u308c\u308b\u3002\u30bf\u30a4\u30de\u30fc\u30921\uff5e2\u6642\u9593\u306b\u30bb\u30c3\u30c8\u3057\u3066\u30b9\u30bf\u30fc\u30c8\u3002<\/li>\n<\/ul>\n<h4 style=\"text-align: left;\">\u2606\u30d6\u30ed\u30c3\u30c6\u30a3\u30f3\u30b0\u2606<\/h4>\n<p style=\"text-align: left;\">Transfer buffer<\/p>\n<p style=\"text-align: left;\">Tris 3.0g<\/p>\n<p style=\"text-align: left;\">Glycine 14.4g<\/p>\n<p style=\"text-align: left;\">SDS 1.0g<\/p>\n<table>\n<tbody>\n<tr>\n<td width=\"397\">\u7279\u7d1aMeOH \u3000\u3000\u3000\u3000200ml<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p style=\"text-align: left;\">Fill up to 1liter(DW)<\/p>\n<ul style=\"text-align: left;\">\n<li>\u30d6\u30ed\u30c3\u30c8\u30e2\u30b8\u30e5\u30fc\u30eb\u306e\u96fb\u6975\u7248\u3092DW\u3067\u6d17\u6d44\u3059\u308b\u3002<\/li>\n<li>\u30b2\u30eb\u3068\u540c\u3058\u30b5\u30a4\u30ba\u306e\u30d6\u30ed\u30c3\u30c6\u30a3\u30f3\u30b0\u30da\u30fc\u30d1\u30fc\u3092\u30b2\u30eb\u306e\u679a\u6570\u00d72\u679a\u6e96\u5099\u3059\u308b\u3002\u30ab\u30c3\u30c8\u3057\u305f\u30da\u30fc\u30d1\u30fc\u306f\u30c8\u30e9\u30f3\u30b9\u30d5\u30a1\u30fc\u30d0\u30c3\u30d5\u30a1\u30fc\u306b\u6d78\u3057\u3066\u304a\u304f\u3002<\/li>\n<li>\u30e1\u30f3\u30d6\u30ec\u30f3\u3092\u540c\u3058\u30b5\u30a4\u30ba\u306b\u30ab\u30c3\u30c8\u3059\u308b\u3002\u30cb\u30c8\u30ed\u30bb\u30eb\u30ed\u30fc\u30b9\u3001\u30ca\u30a4\u30ed\u30f3\u30e1\u30f3\u30d6\u30ec\u30f3\u306fDW\u306b\u3064\u3051\u305f\u5f8c\u3001PVDF\u30e1\u30f3\u30d6\u30ec\u30f3\u306f\u30e1\u30bf\u30ce\u30fc\u30eb\u306b\u3064\u3051\u305f\u5f8c\u306b2\uff5e5\u5206\u9593\u30c8\u30e9\u30f3\u30b9\u30d5\u30a1\u30fc\u30d0\u30c3\u30d5\u30a1\u30fc\u306b\u6d78\u3057\u3066\u304a\u304d\u307e\u3059\u3002\u4e0b\u56f3\u306e\u3088\u3046\u306b\u30c0\u30af\u30ed\u30f3\u30b9\u30dd\u30f3\u30b8(a,f)\u3001\u30d6\u30ed\u30c3\u30c6\u30a3\u30f3\u30b0\u30da\u30fc\u30d1\u30fc(b,e)\u3001\u30b2\u30eb(c)\u3001\u30e1\u30f3\u30d6\u30ec\u30f3(d)\u3092\u91cd\u306d\u308b\u30022\u679a\u306e\u30b2\u30eb\u306e\u30d6\u30ed\u30c3\u30c6\u30a3\u30f3\u30b0\u3092\u540c\u6642\u306b\u884c\u3046\u5834\u5408\u306f\u3001f\u3067\u793a\u3057\u305f2\u679a\u306e\u30c0\u30af\u30ed\u30f3\u30b9\u30dd\u30f3\u30b8\u306e\u9593\u306bb\uff5ee\u3092\u91cd\u306d\u308b\u3002<\/li>\n<\/ul>\n<p style=\"text-align: left;\">\u3000 \u30e1\u30f3\u30d6\u30ec\u30f3\u3068\u30b2\u30eb\u306e\u9593\u306b\u7a7a\u6c17\u304c\u5165\u3089\u306a\u3044\u3088\u3046\u306b\u6ce8\u610f\u3059\u308b\u3002<\/p>\n<hr \/>\n<p><a href=\"https:\/\/www-p.sci.ocha.ac.jp\/bio-shimada-lab\/wp-content\/uploads\/sites\/4\/2015\/08\/clip_image002.jpg\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone size-full wp-image-313\" src=\"https:\/\/www-p.sci.ocha.ac.jp\/bio-shimada-lab\/wp-content\/uploads\/sites\/4\/2015\/08\/clip_image002.jpg\" alt=\"clip_image002\" width=\"291\" height=\"224\" \/><\/a><\/p>\n<p>\uff08\u8d64\uff1a\u967d\u6975\u3001\u9ed2\uff1a\u9670\u6975\uff09<\/p>\n<ul>\n<li>\u9670\u6975\u306e\u4e0a\u306ba\uff5ef\u3092\u306e\u305b\u3001\u30d6\u30ed\u30c3\u30c8\u30e2\u30b8\u30e5\u30fc\u30eb\u3092\u9589\u3058\u30bf\u30d6\u3092\u5965\u307e\u3067\u3057\u3063\u304b\u308a\u3068\u306f\u3081\u8fbc\u3080\u3002<\/li>\n<li>\u30d6\u30ed\u30c3\u30c8\u30e2\u30b8\u30e5\u30fc\u30eb\u3092300\uff5e350ml\u306e\u30c8\u30e9\u30f3\u30b9\u30d5\u30a1\u30fc\u30d0\u30c3\u30d5\u30a1\u30fc\u3067\u6e80\u305f\u3059\u3002<\/li>\n<li>\u30d6\u30ed\u30c3\u30c8\u30e2\u30b8\u30e5\u30fc\u30eb\u3092\u4e0b\u90e8\u30d0\u30c3\u30d5\u30a1\u30fc\u30bf\u30f3\u30af\u306b\u8a2d\u7f6e\u3059\u308b\u3002\u4e0b\u90e8\u30d0\u30c3\u30d5\u30a1\u30fc\u30bf\u30f3\u30af\u30922\uff5e1.7L\u306e\u5ba4\u6e29\u3082\u3057\u304f\u306f4\u2103\u306b\u51b7\u5374\u3057\u305fDW\u3067\u6e80\u305f\u3059\u3002<\/li>\n<li>\u5b9a\u96fb\u6d41300\uff5e400mA\u30671\uff5e2\u6642\u9593\u884c\u3046\u3002<\/li>\n<\/ul>\n","protected":false},"excerpt":{"rendered":"<p>\u30bf\u30f3\u30d1\u30af\u8cea\u62bd\u51fa\u30d7\u30ed\u30c8\u30b3\u30eb \u30fb\u30fb\u30fb\u6e96\u5099\u6bb5\u968e\u30fb\u30fb\u30fb \u85fb\u4f53\u306f\u6c34\u6c17\u3092\u53d6\u3063\u3066\uff0c\u30b3\u30cb\u30ab\u30eb\u30c1\u30e5\u30fc\u30d6\u306b\u3044\u308c\uff0c\u6db2\u4f53\u7a92\u7d20\u3067\u77ac\u9593\u51b7\u51cd\u3057\uff0c-80\u3067\u4fdd\u5b58 \u53ef\u6eb6\u53ef\u30d0\u30c3\u30d5\u30a1\u30fc\u3000Triton-X, Tween 20\u3092\u6e96\u5099 \u30002.2\uff05TritonX-10 &hellip; <a href=\"https:\/\/www-p.sci.ocha.ac.jp\/bio-shimada-lab\/ex_protocol\/pr_06protein.html\">\u7d9a\u304d\u3092\u8aad\u3080 <span class=\"meta-nav\">&rarr;<\/span><\/a><\/p>\n","protected":false},"author":18,"featured_media":0,"parent":94,"menu_order":6,"comment_status":"closed","ping_status":"closed","template":"","meta":{"footnotes":""},"class_list":["post-151","page","type-page","status-publish","hentry"],"aioseo_notices":[],"_links":{"self":[{"href":"https:\/\/www-p.sci.ocha.ac.jp\/bio-shimada-lab\/wp-json\/wp\/v2\/pages\/151","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www-p.sci.ocha.ac.jp\/bio-shimada-lab\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/www-p.sci.ocha.ac.jp\/bio-shimada-lab\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/www-p.sci.ocha.ac.jp\/bio-shimada-lab\/wp-json\/wp\/v2\/users\/18"}],"replies":[{"embeddable":true,"href":"https:\/\/www-p.sci.ocha.ac.jp\/bio-shimada-lab\/wp-json\/wp\/v2\/comments?post=151"}],"version-history":[{"count":14,"href":"https:\/\/www-p.sci.ocha.ac.jp\/bio-shimada-lab\/wp-json\/wp\/v2\/pages\/151\/revisions"}],"predecessor-version":[{"id":352,"href":"https:\/\/www-p.sci.ocha.ac.jp\/bio-shimada-lab\/wp-json\/wp\/v2\/pages\/151\/revisions\/352"}],"up":[{"embeddable":true,"href":"https:\/\/www-p.sci.ocha.ac.jp\/bio-shimada-lab\/wp-json\/wp\/v2\/pages\/94"}],"wp:attachment":[{"href":"https:\/\/www-p.sci.ocha.ac.jp\/bio-shimada-lab\/wp-json\/wp\/v2\/media?parent=151"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}