{"id":4816,"date":"2012-08-10T15:06:38","date_gmt":"2012-08-10T06:06:38","guid":{"rendered":"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/?p=4816"},"modified":"2023-05-15T17:24:06","modified_gmt":"2023-05-15T08:24:06","slug":"maldi-tof%e3%81%ab%e3%82%88%e3%82%8bpmf%e8%a7%a3%e6%9e%90%e3%81%ae%e4%be%8b-2","status":"publish","type":"post","link":"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/?p=4816","title":{"rendered":"MALDI-TOF\u306b\u3088\u308bPMF\u89e3\u6790\u306e\u4f8b 2"},"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 MassFingerprinting; PMF)\u305d\u306e2\u3002\u4e09\u65e5\u76ee\u3067\u3059\u3002<\/p>\n<p>Trypsin\u6d88\u5316\u5f8c\u306e\u30da\u30d7\u30c1\u30c9\u3092\u8131\u5869\u30fb\u6fc3\u7e2e\u3057\u8cea\u91cf\u5206\u6790\u3092\u884c\u3044\u3001\u5143\u306e\u30bf\u30f3\u30d1\u30af\u3092\u540c\u5b9a\u3057\u307e\u3059\u3002<!--more--><\/p>\n<p><u>\u8a66\u85ac\u985e<\/u><\/p>\n<ul>\n<li>\u30da\u30d7\u30c1\u30c9\u6e2c\u5b9a\u7528\u30de\u30c8\u30ea\u30c3\u30af\u30b9 \u03b1-Cyano-4-hydroxycinnamic acid(HCAA) Sigma-Aldrich #C2020 (\u30de\u30c8\u30ea\u30c3\u30af\u30b9\u306f\u65bd\u8a2d\u3067\u5b9f\u8cbb\u3067\u304a\u5206\u3051\u3057\u3066\u3044\u307e\u3059)<\/li>\n<li>\u30ad\u30e3\u30ea\u30d6\u30ec\u30fc\u30b7\u30e7\u30f3\u30b9\u30bf\u30f3\u30c0\u30fc\u30c9<br \/>\nPeptide calibration standard Bruker <a href=\"http:\/\/www.bruker.jp\/daltonics\/imgs\/pdf\/NEW_PL_for_CARE_Products_201108.pdf\">#206195<\/a><\/li>\n<li>\u30c8\u30ea\u30d5\u30eb\u30aa\u30ed\u9162\u9178<br \/>\nTrifluoroacetic Acid \u548c\u5149 206-10736<\/li>\n<li>\u30a2\u30bb\u30c8\u30cb\u30c8\u30ea\u30eb<br \/>\nAcetonitrile \u548c\u5149 019-08631<\/li>\n<\/ul>\n<p><u>\u5668\u5177\u7b49<\/u><\/p>\n<ul>\n<li>ZipTip C18 \u30df\u30ea\u30dd\u30a2 <a href=\"http:\/\/www.millipore.com\/catalogue\/module\/c5737\">ZTC18S008<\/a>\n<\/li>\n<p><a href=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/IMG_1916.jpg\"><img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/IMG_1916-150x150.jpg\" alt=\"\" title=\"IMG_1916\" width=\"150\" height=\"150\" class=\"alignnone size-thumbnail wp-image-4832\" \/><\/a><\/p>\n<li>MTP 384 target plate ground steel TF Bruker <a href=\"http:\/\/www.bruker.jp\/daltonics\/imgs\/pdf\/NEW_PL_for_CARE_Products_201108.pdf\">#209519<\/a><\/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>\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>\u9175\u7d20\u53cd\u5fdc\u5f8c\u306e\u62bd\u51fa\u3068\u6fc3\u7e2e<\/u><\/p>\n<ol>\n<li>\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30bf\u30fc\u304b\u3089\u53d6\u308a\u51fa\u3057\u3001\u8efd\u304fflash\u3057\u3066\u304b\u3089\u62bd\u51fa\u6db2\u309250\u03bcl\u52a0\u3048\u300130\u5206\u9593\u30b7\u30a7\u30fc\u30ab\u30fc\u3067\u632f\u3068\u3046\u3002<\/li>\n<p>\u25cf \u62bd\u51fa\u6db2(\u8981\u6642\u8abf\u6574)<br \/>\n250\u03bcl ACN<br \/>\n 25\u03bcl TFA<br \/>\n225\u03bcl \u30df\u30eaQ\u6c34<\/p>\n<li>\u62bd\u51fa\u6db2\u3092\u3055\u3089\u306b50\u03bcl\u52a0\u3048\u300130\u5206\u9593\u30b7\u30a7\u30fc\u30ab\u30fc\u3067\u632f\u3068\u3046\u3002<\/li>\n<li>\u4e0a\u6e05\u3092\u5225\u306e\u30c1\u30e5\u30fc\u30d6\u306b\u56de\u53ce<\/li>\n<li>\u9060\u5fc3\u30a8\u30d0\u30dd\u30ec\u30fc\u30bf\u30fc\u306710\u03bcl\u7a0b\u5ea6\u306b\u306a\u308b\u307e\u3067\u6fc3\u7e2e\u3059\u308b\u3002<\/li>\n<p>3\u968e\u5171\u540c\u5229\u7528\u5b9f\u9a13\u5ba4\u306eCVE-2000\u3067\u7d0445\u5206\u304f\u3089\u3044\u3002\u98db\u3070\u3057\u3059\u304e\u3066\u3082\u30de\u30ba\u3044\u306e\u3067\u300130\u5206\u3092\u8d85\u3048\u305f\u3042\u305f\u308a\u304b\u308910\u5206\u6bce\u306b\u6a5f\u68b0\u3092\u6b62\u3081\u3066\u91cf\u3092\u78ba\u8a8d\u3057\u305f\u65b9\u304c\u7121\u96e3\u3067\u3059(\u4e00\u5ea6\u30ab\u30e9\u30ab\u30e9\u306b\u3057\u3066\u3057\u307e\u3044\u307e\u3057\u305f\u2026)\u3002\n<\/ol>\n<p><u>\u8131\u5869<\/u><\/p>\n<ol start=5>\n<li>\u8a66\u85ac\u306e\u8abf\u6574<\/li>\n<p>\u25cf \u524d\u51e6\u7406\u6db2(100% TFA)<br \/>\n500\u03bcl ACN<br \/>\n\u25cf \u5e73\u8861\u5316\u6db2(0.1% TFA)\u3000100\u03bcl\u6b8b\u3057\u3066\u6d17\u6d44\u6db2\u306b<br \/>\n1\u03bcl TFA<br \/>\n999\u03bcl \u30df\u30eaQ\u6c34<br \/>\n\u25cf \u6d17\u6d44\u6db2(0.1% TFA)<br \/>\n\u5e73\u8861\u5316\u6db2\u3092\u6d41\u7528\u3002\u30b5\u30f3\u30d7\u30eb\u5f53\u308a100\u03bcl\u306b\u5206\u6ce8\u3057\u3066\u304a\u304f\u3068\u826f\u3044<br \/>\n\u25cf \u6eb6\u51fa\u6db2(50% ACN, 0.1% TFA)<br \/>\n1\u03bcl   TFA<br \/>\n500\u03bcl ACN<br \/>\n499\u03bcl \u30df\u30eaQ\u6c34<br \/>\n\u6eb6\u51fa\u6db2\u3082\u30b5\u30f3\u30d7\u30eb\u5f53\u308a100\u03bcl\u306b\u5206\u6ce8\u3057\u3066\u304a\u304f\u3068\u826f\u3044<\/p>\n<li>ZipTip\u3092\u30d4\u30da\u30c3\u30c8\u306b\u88c5\u7740\u3057\u300110\u03bcl\u306b\u30bb\u30c3\u30c8<\/li>\n<li>\u524d\u51e6\u7406\u6db2\u3092\u3086\u3063\u304f\u308a\u3086\u3063\u304f\u308a\u5438\u5f15\u3057\u3001\u5ec3\u6db2\u5165\u308c\u306b\u3086\u3063\u304f\u308a\u3086\u3063\u304f\u308a\u6392\u51fa\u3002\u3053\u306e\u64cd\u4f5c\u30925\u56de\u7e70\u308a\u8fd4\u3059\u3002<\/li>\n<li>\u5e73\u8861\u5316\u6db2\u3092\u3086\u3063\u304f\u308a\u3086\u3063\u304f\u308a\u5438\u5f15\u3057\u3001\u5ec3\u6db2\u5165\u308c\u306b\u3086\u3063\u304f\u308a\u3086\u3063\u304f\u308a\u6392\u51fa\u3002\u3053\u306e\u64cd\u4f5c\u30925\u56de\u7e70\u308a\u8fd4\u3059\u3002<\/li>\n<li>\u30b5\u30f3\u30d7\u30eb\u3092\u3086\u3063\u304f\u308a\u3086\u3063\u304f\u308a5\u56de\u30d4\u30da\u30c3\u30c6\u30a3\u30f3\u30b0\u3057\u3066\u3001\u30ec\u30b8\u30f3\u306b\u5438\u7740\u3055\u305b\u308b<\/li>\n<li>\u6d17\u6d44\u6db2\u3092\u3086\u3063\u304f\u308a\u3086\u3063\u304f\u308a\u5438\u5f15\u3057\u3001\u5ec3\u6db2\u5165\u308c\u306b\u3086\u3063\u304f\u308a\u3086\u3063\u304f\u308a\u6392\u51fa\u3002\u3053\u306e\u64cd\u4f5c\u30925\u56de\u7e70\u308a\u8fd4\u3059\u3002<\/li>\n<li>\u30d4\u30da\u30c3\u30c8\u30923\u03bcl\u306b\u30bb\u30c3\u30c8<\/li>\n<li>\u6eb6\u51fa\u6db2\u3092\u3086\u3063\u304f\u308a\u3086\u3063\u304f\u308a\u5438\u5f15\u3057\u3001\u5225\u306e\u65b0\u3057\u3044\u30c1\u30e5\u30fc\u30d6\u306b\u6392\u51fa\u3002\u305d\u306e\u307e\u307e5\u56de\u7a0b\u5ea6\u30d4\u30da\u30c3\u30c6\u30a3\u30f3\u30b0\u3002<\/li>\n<p>\u6eb6\u51fa\u6db2\u304c\u7121\u304f\u306a\u3063\u3066\u3057\u307e\u3063\u305f\u5834\u5408\u306f\u3001\u614c\u3066\u305a\u306b\u3055\u3089\u306b3\u03bcl\u8ffd\u52a0\u3057\u3066\u3082\u5927\u4e08\u592b\u3002<br \/>\n\u3059\u3050\u6e2c\u5b9a\u3057\u306a\u3044\u306e\u3067\u3042\u308c\u30704\u2103\u4fdd\u5b58\u306e\u3053\u3068(\u5fae\u91cf\u306a\u306e\u3067\u3067\u304d\u308b\u3060\u3051\u65e9\u304f\u6e2c\u5b9a\u3057\u305f\u65b9\u304c\u826f\u3044\u3067\u3059)\u3002\n<\/ol>\n<p><u>\u6e2c\u5b9a<\/u><\/p>\n<ol start=13>\n<li>\u30ad\u30e3\u30ea\u30d6\u30ec\u30fc\u30b7\u30e7\u30f3\u30b9\u30bf\u30f3\u30c0\u30fc\u30c9\u3092\u6eb6\u304b\u3057\u3066\u304a\u304f\u3002<\/li>\n<li>\u30de\u30c8\u30ea\u30c3\u30af\u30b9\u6eb6\u6db2\u306e\u8abf\u6574<\/li>\n[TA]\n400\u03bcl 0.1% TFA<br \/>\n200\u03bcl ACN<\/p>\n<p>\u30de\u30a4\u30af\u30ed\u30b9\u30d1\u30fc\u30c6\u30eb1\u676f\u5206\u306eHCAA\u3092\u30c1\u30e5\u30fc\u30d6\u306b\u53d6\u308a\u3001500\u03bcl\u306eTA\u3092\u52a0\u3048\u3066\u30dc\u30eb\u30c6\u30c3\u30af\u30b9(\u98fd\u548c\u6eb6\u6db2\u306e\u305f\u3081\u5b8c\u5168\u306b\u306f\u6eb6\u3051\u306a\u3044)\u3002<br \/>\n\u30c1\u30d3\u30bf\u30f3\u7b49\u3067Flash\u3057\u3066\u4e0a\u6e05\u3092\u5225\u306e\u30c1\u30e5\u30fc\u30d6\u306b\u79fb\u3059\u3002<\/p>\n<li>\u30b5\u30f3\u30d7\u30eb\u304a\u3088\u3073\u30ad\u30e3\u30ea\u30d6\u30ec\u30fc\u30b7\u30e7\u30f3\u30b9\u30bf\u30f3\u30c0\u30fc\u30c9\u3092\u305d\u308c\u305e\u308c1:4\u306e\u6bd4\u7387\u3067\u30de\u30c8\u30ea\u30c3\u30af\u30b9\u6eb6\u6db2\u3068\u6df7\u5408\u3057\u305f\u3082\u306e\u3092\u8abf\u6574\u3002(\u30d4\u30da\u30c3\u30c6\u30a3\u30f3\u30b0\u306b\u3088\u308a\u6df7\u5408)<\/li>\n<li>1\u03bcl\u305a\u3064\u30bf\u30fc\u30b2\u30c3\u30c8\u30d7\u30ec\u30fc\u30c8\u306b\u8f09\u305b\u3066\u98a8\u4e7e\u5f8cAutoflex\u3067\u6e2c\u5b9a<\/li>\n<p><a href=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/P1010824.jpg\"><img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/P1010824-300x225.jpg\" alt=\"\" title=\"P1010824\" width=\"150\" height=\"112\" class=\"alignnone size-medium wp-image-4856\" srcset=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/P1010824-300x225.jpg 300w, https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/P1010824-1024x768.jpg 1024w\" sizes=\"auto, (max-width: 150px) 100vw, 150px\" \/><\/a><a href=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/P1010825.jpg\"><img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/P1010825-300x225.jpg\" alt=\"\" title=\"P1010825\" width=\"150\" height=\"112\" class=\"alignnone size-medium wp-image-4857\" srcset=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/P1010825-300x225.jpg 300w, https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/P1010825-1024x768.jpg 1024w\" sizes=\"auto, (max-width: 150px) 100vw, 150px\" \/><\/a>\n<\/ol>\n<p><u>\u7d50\u679c<\/u><br \/>\n\u4eca\u56de\u8abf\u6574\u3057\u305fBSA\u306e\u7d50\u679c\u306f\u3053\u306e\u3088\u3046\u306a\u5177\u5408\u3067\u3057\u305f\u3002<br \/>\n<a href=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/Image81.png\"><img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/Image81-300x296.png\" alt=\"\" title=\"Image81\" width=\"300\" height=\"296\" class=\"alignnone size-medium wp-image-4822\" srcset=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/Image81-300x296.png 300w, https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/Image81.png 974w\" sizes=\"auto, (max-width: 300px) 100vw, 300px\" \/><\/a><br \/>\nMascot\u30b5\u30fc\u30c1\u306e\u7d50\u679c\u3001\u7121\u4e8b\u306b\u540c\u5b9a\u3055\u308c\u3066\u3044\u308b\u3088\u3046\u3067\u3059\u3002<br \/>\n\u307b\u3068\u3093\u3069\u624b\u888b\u3092\u7740\u7528\u3057\u306a\u304b\u3063\u305f\u308a\u306a\u3069\u3001\u304b\u306a\u308a\u30e9\u30d5\u306b\u5b9f\u9a13\u3057\u305f\u3053\u3068\u3082\u3042\u308a\u3001Mascot\u306e\u30b9\u30b3\u30a2\u306f\u3053\u3093\u306a\u3082\u306e\u304b\u306a\u3063\u3066\u611f\u3058\u3067\u3059\u3002<\/p>\n<p>\u3061\u306a\u307f\u306b\u3001ZipTip\u3092\u4f7f\u308f\u306a\u3044\u3067\u3001EASY-nLC\u3092\u4f7f\u3063\u305f<a href=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/?p=4728\">LC-MALDI<\/a>\u3067\u540c\u3058\u30b5\u30f3\u30d7\u30eb\u3092\u6e2c\u5b9a\u3059\u308b\u3068\u3001\u3053\u3093\u306a\u611f\u3058\u306b\u3002<br \/>\n<a href=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/Image23.png\"><img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/Image23-275x300.png\" alt=\"\" title=\"Image23\" width=\"275\" height=\"300\" class=\"alignnone size-medium wp-image-4825\" srcset=\"https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/Image23-275x300.png 275w, https:\/\/gene4.knit.kagoshima-u.ac.jp\/grc\/wp-content\/uploads\/2012\/08\/Image23.png 907w\" sizes=\"auto, (max-width: 275px) 100vw, 275px\" \/><\/a><br \/>\n\u30b9\u30b3\u30a2\u306fZipTip\u306e67\u306b\u5bfe\u3057\u3066172\u307e\u3067\u4e0a\u304c\u308a\u307e\u3059\u3002<\/p>\n<p>\u4ee5\u4e0a\u304cMALDI-TOF\u306b\u3088\u308bPMF\u89e3\u6790\u306e\u6d41\u308c\u3068\u306a\u308a\u307e\u3059\u3002<br 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