{"id":4763,"date":"2012-08-10T15:06:33","date_gmt":"2012-08-10T06:06:33","guid":{"rendered":"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/?p=4763"},"modified":"2012-11-05T17:38:06","modified_gmt":"2012-11-05T08:38:06","slug":"maldi-tof%e3%81%ab%e3%82%88%e3%82%8bpmf%e8%a7%a3%e6%9e%90%e3%81%ae%e4%be%8b-1","status":"publish","type":"post","link":"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/?p=4763","title":{"rendered":"MALDI-TOF\u306b\u3088\u308bPMF\u89e3\u6790\u306e\u4f8b 1"},"content":{"rendered":"<p>\u8cea\u91cf\u5206\u6790\u88c5\u7f6eAutoflex Speed\u306b\u3088\u308b\u30da\u30d7\u30c1\u30c9\u30fb\u30de\u30b9\u30fb\u30d5\u30a3\u30f3\u30ac\u30fc\u30d7\u30ea\u30f3\u30c6\u30a3\u30f3\u30b0(Peptide Mass Fingerprinting; PMF)\u306e\u4e00\u4f8b\u3067\u3059\u3002\u907a\u4f1d\u5b50\u5b9f\u9a13\u65bd\u8a2d\u306e\u6a5f\u6750\u3092\u4f7f\u3063\u3066\u5b9f\u65bd\u3057\u3066\u307f\u307e\u3057\u305f\u3002\u53c2\u8003\u306b\u306a\u308c\u3070\u5e78\u3044\u3067\u3059\u3002<\/p>\n<p>\u5b9f\u9a13\u306f\u6982\u306d3\u65e5\u9593\u304b\u304b\u308a\u307e\u3059\u3002<br \/>\n\u521d\u65e5\u306f\u96fb\u6c17\u6cf3\u52d5\u3068\u30b2\u30eb\u67d3\u8272\u3002\u4e8c\u65e5\u76ee\u306f\u30b2\u30eb\u304b\u3089\u306e\u30d0\u30f3\u30c9\u306e\u5207\u308a\u51fa\u3057\u304b\u3089\u30b2\u30eb\u5185\u9175\u7d20\u6d88\u5316\u307e\u3067\u3002\u4e09\u65e5\u76ee\u306f\u9175\u7d20\u6d88\u5316\u3055\u308c\u305f\u30da\u30d7\u30c1\u30c9\u306e\u8131\u5869\u30fb\u6fc3\u7e2e\u3068\u8cea\u91cf\u5206\u6790\u3068\u3044\u3046\u30b9\u30b1\u30b8\u30e5\u30fc\u30eb\u3067\u3059\u3002<br \/>\n\u4eca\u56de\u306f\u4e8c\u65e5\u76ee\u306e\u30d0\u30f3\u30c9\u5207\u308a\u51fa\u3057\u304b\u3089\u7d39\u4ecb\u3057\u3066\u3044\u304d\u307e\u3059\u3002<br \/>\n<!--more--><br \/>\n<img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/IMG_1908-150x150.jpg\" alt=\"\" title=\"IMG_1908\" width=\"150\" height=\"150\" class=\"alignleft size-thumbnail wp-image-4796\" \/><br \/>\n\u4eca\u56de\u306f\u5c02\u4efb\u6559\u5b98\u5b9f\u9a13\u5ba4\u306e\u30d5\u30ea\u30fc\u30b6\u30fc\u306b\u7720\u3063\u3066\u3044\u305f(\u671f\u9650\u5207\u308c\u306e)BSA\u3092\u4f7f\u3063\u3066\u3001\u3069\u3061\u3089\u304b\u3068\u3044\u3046\u3068\u304b\u306a\u308a\u30e9\u30d5\u306b\u5b9f\u9a13\u3057\u3066\u3044\u307e\u3059\u3002\u3053\u306eBSA\u306f\u30d7\u30ed\u30c6\u30aa\u30df\u30af\u30b9\u7528\u306e\u30de\u30fc\u30ab\u30fc\u8a66\u85ac\u3067\u306f\u306a\u304f\u3001\u304a\u305d\u3089\u304f\u904e\u53bb\u306b\u30b5\u30b6\u30f3\u30cf\u30a4\u30d6\u30ea\u7b49\u306e\u30d6\u30ed\u30c3\u30ad\u30f3\u30b0\u8a66\u85ac\u3068\u3057\u3066\u4f7f\u3063\u3066\u3044\u305f\u3082\u306e\u3067\u3001\u305a\u3063\u3068\u5fd8\u308c\u3089\u308c\u3066\u3044\u305f\u3082\u306e\u3067\u3059\u3002\u679c\u305f\u3057\u3066\u7121\u4e8b\u306bBSA\u3068\u540c\u5b9a\u3067\u304d\u308b\u3067\u3057\u3087\u3046\u304b\u3002<\/p>\n<p><u>\u30b9\u30c8\u30c3\u30af\u8abf\u6574\u8a66\u85ac\u7b49<\/u><br \/>\n\u30b9\u30c8\u30c3\u30af\u8a66\u85ac\u4ee5\u5916\u306e\u8a66\u85ac\u985e\u306f\u3059\u3079\u3066\u8981\u6642\u8abf\u6574\u3002\u307e\u305f\u3001\u6c34\u306f\u3059\u3079\u3066<a href=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/?page_id=2918\">\u30df\u30eaQ\u6c34<\/a>\u3092\u7528\u3044\u308b\u3053\u3068(\u96fb\u6c17\u6cf3\u52d5\u3082)\u3002<\/p>\n<ul>\n<li>1M NH<sub>4<\/sub>HCO<sub>3<\/sub>(\u70ad\u9178\u6c34\u7d20\u30a2\u30f3\u30e2\u30cb\u30a6\u30e0)<\/li>\n<p>Ammonium Hydrogen Carbonate \u30ca\u30ab\u30e9\u30a4\u30c6\u30b9\u30af 1066-33-7<br \/>\n\t0.79g\u306eNH<sub>4<\/sub>HCO<sub>3<\/sub>\u309210ml\u306e\u30df\u30eaQ\u6c34\u306b\u6eb6\u304b\u3059\u3002<\/p>\n<li>1M \u30b8\u30c1\u30aa\u30b9\u30ec\u30a4\u30c8\u30fc\u30eb(DTT)<\/li>\n<p>Dithiothreitol \u30ca\u30ab\u30e9\u30a4\u30c6\u30b9\u30af 27565-41-9<br \/>\n\t154.2mg\u306e\u30921ml\u306e\u30df\u30eaQ\u6c34\u306b\u6eb6\u304b\u3059\u3002(\u30de\u30a4\u30af\u30ed\u30c1\u30e5\u30fc\u30d6\u3054\u3068\u5929\u79e4\u3067\u79e4\u91cf\u3057\u3001\u540c\u3058\u6bd4\u7387\u306e\u6c34\u3092\u52a0\u3048\u308c\u3070\u826f\u3044\u300250\u03bcl\u306b\u5206\u6ce8\u3057\u51b7\u51cd\u4fdd\u5b58)<\/p>\n<li>Trypsin\u30b9\u30c8\u30c3\u30af\u6eb6\u6db2<\/li>\n<p>\tPromega Sequence Grade Modified Trypsin V511A<br \/>\n\u6dfb\u4ed8Buffer200\u03bcl\u3067\u30d0\u30a4\u30a2\u30eb1\u672c\u5206\u3092\u6eb6\u89e3\u300210\u03bcl\u305a\u3064\u306b\u5206\u6ce8\u3057-80\u2103\u4fdd\u5b58\u3002\n<\/ul>\n<p><u>\u5668\u5177\u7b49<\/u><\/p>\n<ul>\n<li>\u30e1\u30b9(\u30c7\u30a3\u30b9\u30dd\u30fc\u30b6\u30d6\u30eb)<\/li>\n<li>\u30d4\u30f3\u30bb\u30c3\u30c8<\/li>\n<li>\u624b\u888b(\u30c7\u30a3\u30b9\u30dd\u30fc\u30b6\u30d6\u30eb)<\/li>\n<li>\u9060\u5fc3\u30a8\u30d0\u30dd\u30ec\u30fc\u30bf <a href=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/?page_id=3523\">CVE-2000<\/a> or <a href=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/?page_id=2830\">VC-96N<\/a><\/li>\n<li>\u30b7\u30a7\u30fc\u30ab\u30fc Iwaki TWIN3-28N (3\u968e\u5eca\u4e0b\u306b\u3042\u308a\u307e\u3059)<\/li>\n<p><a href=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/IMG_1909.jpg\"><img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/IMG_1909-150x150.jpg\" alt=\"\" title=\"IMG_1909\" width=\"150\" height=\"150\" class=\"alignnone size-thumbnail wp-image-4799\" \/><\/a><\/p>\n<li>\u30d6\u30ed\u30c3\u30af\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30bf\u30fc TEITEC DTU-1B (3\u968e\u5171\u540c\u5229\u7528\u5b9f\u9a13\u5ba4\u306b\u3042\u308a\u307e\u3059)<\/li>\n<p><a href=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/IMG_1912.jpg\"><img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/IMG_1912-150x150.jpg\" alt=\"\" title=\"IMG_1912\" width=\"150\" height=\"150\" class=\"alignnone size-thumbnail wp-image-4800\" \/><\/a><\/p>\n<li>\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30bf\u30fc <a href=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/?page_id=2861\">IC600<\/a> or <a href=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/?page_id=3568\">HB-80<\/a><\/li>\n<li>\u305d\u306e\u4ed6\u3001\u30d4\u30da\u30c3\u30c8\u304a\u3088\u3073\u30c1\u30d3\u30bf\u30f3\u306a\u3069\u30de\u30a4\u30af\u30ed\u30c1\u30e5\u30fc\u30d6\u7528\u9060\u5fc3\u6a5f<\/li>\n<\/ul>\n<p><u>\u30b2\u30eb\u304b\u3089\u306e\u5207\u308a\u51fa\u3057<\/u><\/p>\n<ol>\n<li>\u30a6\u30b7\u8840\u6e05\u30a2\u30eb\u30d6\u30df\u30f3(\u30ca\u30ab\u30e9\u30a4, F-V) 5pM \u309210% SDS-PAGE\u3067\u5206\u96e2\u3057CBB\u67d3\u8272\u3057\u3066\u304a\u304f\u3002<\/li>\n<p><img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/IMG_1728-300x196.jpg\" alt=\"\" title=\"IMG_1728\" width=\"250\" height=\"163\" class=\"alignnone size-medium wp-image-4768\" srcset=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/IMG_1728-300x196.jpg 300w, https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/IMG_1728-1024x672.jpg 1024w, https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/IMG_1728.jpg 1828w\" sizes=\"auto, (max-width: 250px) 100vw, 250px\" \/><\/p>\n<li>66kDa\u4ed8\u8fd1\u306e\u30d0\u30f3\u30c9\u3092\u30e1\u30b9\u3067\u5207\u308a\u51fa\u3057\u30014,5\u7247\u7a0b\u5ea6\u306b\u7d30\u5207\u3057\u3066\u30de\u30a4\u30af\u30ed\u30c1\u30e5\u30fc\u30d6\u306b\u5165\u308c\u308b\u3002<\/li>\n<p><a href=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/IMG_1729.jpg\"><img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/IMG_1729-300x203.jpg\" alt=\"\" title=\"IMG_1729\" width=\"250\" height=\"169\" class=\"alignnone size-medium wp-image-4770\" srcset=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/IMG_1729-300x203.jpg 300w, https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/IMG_1729-1024x693.jpg 1024w, https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/IMG_1729.jpg 1500w\" sizes=\"auto, (max-width: 250px) 100vw, 250px\" \/><\/a><br \/>\n\u30e1\u30b9\u3084\u30d4\u30f3\u30bb\u30c3\u30c8\u306f\u305d\u306e\u90fd\u5ea670%\u30a8\u30bf\u30ce\u30fc\u30eb\u3068\u30ad\u30e0\u30ef\u30a4\u30d7\u3067\u304d\u308c\u3044\u306b\u62ed\u3044\u3066\u4f7f\u3046\u3053\u3068\u3002\u5834\u5408\u306b\u3088\u3063\u3066\u306f\u30b5\u30f3\u30d7\u30eb(\u30d0\u30f3\u30c9)\u3054\u3068\u306b\u4ea4\u63db\u3057\u305f\u65b9\u304c\u826f\u3044\u304b\u3082\u3002<\/ol>\n<p><u>\u8131\u8272<\/u><\/p>\n<ol start=3>\n<li>\u30c1\u30e5\u30fc\u30d6\u5f53\u308a100\u03bcl\u306e\u8131\u8272\u6db2\u3092\u52a0\u3048\u30b7\u30a7\u30fc\u30ab\u30fc\u306710\u5206\u9593\u632f\u3068\u3046\u3059\u308b\u3002(\u3053\u306e\u9593\u306b\u30d6\u30ed\u30c3\u30af\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30bf\u30fc\u309256\u2103\u306b\u30bb\u30c3\u30c8)<\/li>\n[\u8131\u8272\u6db2(\u8981\u4e8b\u8abf\u6574)]\n100\u03bcl  1M NH<sub>4<\/sub>HCO<sub>3<\/sub><br \/>\n1.2ml  \u30a2\u30bb\u30c8\u30cb\u30c8\u30ea\u30eb(ACN)<br \/>\n2.7ml  MiliQ\u6c34<\/p>\n<li>\u4e0a\u6e05\u3092\u9664\u53bb\u3002\u30b2\u30eb\u306e\u8272\u304c\u6d88\u3048\u308b\u307e\u3067\u3053\u306e\u64cd\u4f5c\u3092\u7e70\u308a\u8fd4\u3059(2\u301c3\u56de\u7a0b\u5ea6)\u3002<\/li>\n<li>\u4e0a\u6e05\u3092\u9664\u53bb\u3057\u3001150\u03bcl\u306eACN\u3092\u52a0\u3048\u3066\u66f4\u306b2\u5206\u9593\u632f\u3068\u3046\u5f8c\u3001\u4e0a\u6e05\u3092\u9664\u53bb\u3002<\/li>\n<li>\u9060\u5fc3\u30a8\u30d0\u30dd\u30ec\u30fc\u30bf\u30fc\u30675\u5206\u9593\u771f\u7a7a\u4e7e\u71e5\u3002<\/li>\n<\/ol>\n<p><u>\u9084\u5143\u30fb\u30a2\u30eb\u30ad\u30eb\u5316<\/u><\/p>\n<ol start=7>\n<li>\u9084\u5143\u6db2\u3092100\u03bcl\u52a0\u3048\u300156\u2103\u30671\u6642\u9593\u53cd\u5fdc\u3055\u305b\u308b<\/li>\n[\u9084\u5143\u6db2(\u4f7f\u7528\u76f4\u524d\u306b\u6df7\u5408\u8abf\u6574)]\n10\u03bcl  1M DTT<br \/>\n25\u03bcl  1M NH<sub>4<\/sub>HCO<sub>3<\/sub><br \/>\n965\u03bcl \u30df\u30eaQ\u6c34<\/p>\n<li>\u4e0a\u6e05\u3092\u9664\u53bb\u3059\u308b\u3002(\u30c1\u30d3\u30bf\u30f3\u7b49\u3067Flash\u3057\u3066\u96c6\u3081\u3066\u5b8c\u5168\u306b\u9664\u304f\u3053\u3068)<\/li>\n<li>\u6d17\u6d44\u6db2\u3092100\u03bcl\u52a0\u3048\u3001\u30b7\u30a7\u30fc\u30ab\u30fc\u306710\u5206\u9593\u632f\u3068\u3046\u3002<\/li>\n[\u6d17\u6d44\u6db2(\u4f7f\u7528\u76f4\u524d\u306b\u6df7\u5408\u8abf\u6574\u30001ml\u306e\u6eb6\u6db2\u30923\u672c\u7528\u610f\u3059\u308b\u3068\u826f\u3044)]\n25\u03bcl  1M NH<sub>4<\/sub>HCO<sub>3<\/sub><br \/>\n975\u03bcl \u30df\u30eaQ\u6c34<\/p>\n<li>\u4e0a\u6e05\u3092\u9664\u53bb\u3059\u308b\u3002<\/li>\n<li>\u30a2\u30eb\u30ad\u30eb\u5316\u6db2\u3092100\u03bcl\u52a0\u3048\u3001\u30b7\u30a7\u30fc\u30ab\u30fc\u306745\u5206\u9593\u632f\u3068\u3046\u3002\u3053\u306e\u3068\u304d\u3001\u30b7\u30a7\u30fc\u30ab\u30fc\u3092\u4e38\u3054\u3068\u6bb5\u30dc\u30fc\u30eb\u7bb1\u7b49\u3067\u8986\u3063\u3066\u906e\u5149\u3059\u308b\u3002\u5834\u5408\u306b\u3088\u3063\u3066\u306f\u30a2\u30eb\u30df\u30db\u30a4\u30eb\u3067\u30b7\u30a7\u30fc\u30ab\u30fc\u3054\u3068\u8986\u3063\u3066\u3057\u307e\u3046\u3002<\/li>\n[\u30a2\u30eb\u30ad\u30eb\u5316\u6db2(\u4f7f\u7528\u76f4\u524d\u306b\u6df7\u5408\u8abf\u6574)]\n0.01g \u30e8\u30fc\u30c9\u30a2\u30bb\u30c8\u30a2\u30df\u30c9<br \/>\n1ml   \u6d17\u6d44\u6db2<br \/>\n<u>\u30e8\u30fc\u30c9\u30a2\u30bb\u30c8\u30a2\u30df\u30c9\u306f\u6709\u6bd2\u306a\u305f\u3081\u624b\u888b\u3092\u7740\u7528\u306e\u3053\u3068\u3002\u5929\u79e4\u3067\u30c1\u30e5\u30fc\u30d6\u306b\u76f4\u63a5\u79e4\u91cf\u3057\u3001\u540c\u3058\u6bd4\u7387\u306e\u6d17\u6d44\u6db2\u3067\u6eb6\u304b\u3059\u3002<\/u><\/p>\n<li>\u4e0a\u6e05\u3092\u5b8c\u5168\u306b\u9664\u53bb\u3002<\/li>\n<li>\u6d17\u6d44\u6db2\u3092100\u03bcl\u52a0\u3048\u3001\u30b7\u30a7\u30fc\u30ab\u30fc\u306710\u5206\u9593\u632f\u3068\u3046\u3002<\/li>\n<li>\u4e0a\u6e05\u3092\u5b8c\u5168\u306b\u9664\u53bb\u3002<\/li>\n<\/ol>\n<p><u>\u30b2\u30eb\u5185\u6d88\u5316<\/u><\/p>\n<ol start=15>\n<li>\u8131\u6c34\u6db2\u3092200\u03bcl\u52a0\u3048\u3001\u30b7\u30a7\u30fc\u30ab\u30fc\u306710\u5206\u9593\u632f\u3068\u3046\u3002<\/li>\n[\u8131\u6c34\u6db2(\u4f7f\u7528\u76f4\u524d\u306b\u6df7\u5408\u8abf\u6574)]\n2ml   ACN<br \/>\n100\u03bcl 1M NH<sub>4<\/sub>HCO<sub>3<\/sub><br \/>\n1.9ml 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