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1\u30a6\u30a7\u30b9\u30bf\u30f3\u30d6\u30ed\u30c3\u30c6\u30a3\u30f3\u30b0\u3067\u6297\u539f\u3092\u691c\u51fa\u3067\u304d\u306a\u3044\u3053\u3068\u304c\u3042\u308a\u307e\u3059\u3002\u4e00\u6b21\u6297\u4f53\u3092\u9078\u3076\u3068\u304d\u306f\u3001\u30a2\u30d7\u30ea\u30b1\u30fc\u30b7\u30e7\u30f3\u6b04\u3092\u3057\u3063\u304b\u308a\u78ba\u8a8d\u3057\u3066\u304a\u304d\u307e\u3057\u3087\u3046\u3002\r\n\u7279\u7570\u6027\u691c\u8a3c\u6e08\u307f\u306e\u4e00\u6b21\u6297\u4f53\u3082\r\n\u30a6\u30a7\u30b9\u30bf\u30f3\u30d6\u30ed\u30c3\u30c6\u30a3\u30f3\u30b0\u3067\u7121\u4e8b\u306b\u691c\u51fa\u3067\u304d\u305f\u3051\u308c\u3069\u3001\u679c\u305f\u3057\u3066\u3053\u306e\u30d0\u30f3\u30c9\u306f\u672c\u5f53\u306b\u76ee\u7684\u30bf\u30f3\u30d1\u30af\u8cea\u3060\u308d\u3046\u304b\uff1f\u305d\u3093\u306a\u7591\u554f\u3068\u5411\u304d\u5408\u3063\u305f\u7d4c\u9a13\u304c\u3042\u308b\u65b9\u3082\u3044\u308b\u304b\u3082\u3057\u308c\u307e\u305b\u3093\u3002\u6297\u4f53\u306e\u7279\u7570\u6027\u3084\u6027\u80fd\u306b\u3064\u3044\u3066\u5b9f\u969b\u306b\u691c\u8a3c\u3055\u308c\u305f\u6297\u4f53\u3092\u4f7f\u3044\u305f\u3044\u3068\u3044\u3046\u7814\u7a76\u8005\u306f\u591a\u304f\u3044\u3089\u3063\u3057\u3083\u3044\u307e\u3059\u3002\u3053\u3046\u3044\u3063\u305f\u80cc\u666f\u304b\u3089\u3001\u6297\u4f53\u304c\u6b63\u3057\u304f\u30bf\u30fc\u30b2\u30c3\u30c8\u3068\u7d50\u5408\u3057\u305f\u3053\u3068\u3092\u8a3c\u660e\u3057\u305f\u300c\u691c\u8a3c\u6e08\u307f\u306e\u6297\u4f53\u300d\u3068\u3044\u3046\u3082\u306e\u304c\u5b58\u5728\u3057\u307e\u3059\u3002\r\n\r\n\u30e1\u30fc\u30ab\u30fc\u306b\u3088\u3063\u3066\u547c\u3073\u540d\u306f\u7570\u306a\u308a\u307e\u3059\u304c\u3001\u305f\u3068\u3048\u3070\u3001\u30b5\u30fc\u30e2\u30d5\u30a3\u30c3\u30b7\u30e3\u30fc\u30b5\u30a4\u30a8\u30f3\u30c6\u30a3\u30d5\u30a3\u30c3\u30af\u3067\u306f\u3001\u300cAdvanced Verification\u300d\u3068\u3044\u3046\u30a2\u30a4\u30b3\u30f3\u304c\u3064\u3044\u3066\u3044\u307e\u3059\u3002\u30ce\u30c3\u30af\u30c0\u30a6\u30f3\u30fb\u30ce\u30c3\u30af\u30a2\u30a6\u30c8\u30fb\u4e2d\u548c\u30fbIP\u3068\u8cea\u91cf\u5206\u6790\u6cd5\u306a\u3069\u3001\u898f\u5b9a\u3055\u308c\u305f\u8907\u6570\u306e\u691c\u8a3c\u65b9\u6cd5\u306e\u3046\u3061\u5c11\u306a\u304f\u3068\u30821\u7a2e\u985e\u4ee5\u4e0a\u306e\u691c\u8a3c\u3092\u5b9f\u65bd\u3057\u3066\u3044\u308b\u6297\u4f53\u3067\u3059\u3002\u4f8b\u3048\u3070\u30ce\u30c3\u30af\u30a2\u30a6\u30c8\u691c\u8a3c\u3067\u306f\u3001CRISPR\/Cas9\u3092\u7528\u3044\u3066\u3001\u30bf\u30fc\u30b2\u30c3\u30c8\u30bf\u30f3\u30d1\u30af\u8cea\u306e\u30ce\u30c3\u30af\u30a2\u30a6\u30c8\u682a\u3092\u7528\u3044\u3066\u8a55\u4fa1\u3092\u3057\u3066\u3044\u307e\u3059\u3002\u3082\u3057\u6297\u4f53\u306e\u7279\u7570\u6027\u306b\u4e0d\u5b89\u304c\u3042\u308b\u5834\u5408\u306f\u3001\u3053\u3046\u3044\u3063\u305f\u691c\u8a3c\u6e08\u307f\u306e\u6297\u4f53\u3092\u9078\u629e\u3057\u307e\u3057\u3087\u3046\u3002\r\n\u4e8c\u6b21\u6297\u4f53\u9078\u629e\u6642\u306b\u77e5\u3063\u3066\u304a\u304d\u305f\u3044\u3053\u3068\r\n\u307e\u305a\u3001\u57fa\u672c\u3068\u3057\u3066\u3001\u4e8c\u6b21\u6297\u4f53\u306f\u3001\u4e00\u6b21\u6297\u4f53\u306eHost\uff08\u7531\u6765\u30fb\u514d\u75ab\u52d5\u7269\uff09\u306b\u5fdc\u3058\u3066\u9078\u3073\u307e\u3057\u3087\u3046\u3002\u4f8b\u3048\u3070\u4e00\u6b21\u6297\u4f53\u306eHost\u304cmouse\u3067\u3042\u308c\u3070\u3001\u4e8c\u6b21\u6297\u4f53\u306fAnti-Mouse\u3092\u9078\u629e\u3057\u307e\u3059\u3002\r\n\r\n\u4e8c\u6b21\u6297\u4f53\u3067\u306f\u3001\u4f55\u3089\u304b\u306e\u6a19\u8b58\u304c\u3042\u308b\u3082\u306e\u3092\u7528\u3044\u308b\u3053\u3068\u304c\u4e00\u822c\u7684\u3067\u3059\u3002\u86cd\u5149\u691c\u51fa\u3067\u3042\u308c\u3070\u3001\u86cd\u5149\u6a19\u8b58\u306e\u4e8c\u6b21\u6297\u4f53\u3001\u5316\u5b66\u767a\u5149\u3067\u691c\u51fa\u3059\u308b\u306e\u3067\u3042\u308c\u3070\u3001HRP\u306a\u3069\u306e\u9175\u7d20\u6a19\u8b58\u306e\u3082\u306e\u3092\u7528\u3044\u307e\u3059\u3002\r\n\u307e\u305f\u3001\u66f8\u304b\u308c\u3066\u3044\u306a\u3044\u3053\u3068\u3082\u3042\u308a\u307e\u3059\u304c\u3001\u307b\u3068\u3093\u3069\u306e\u4e8c\u6b21\u6297\u4f53\u306f\u30dd\u30ea\u30af\u30ed\u30fc\u30ca\u30eb\u6297\u4f53\u3067\u3059\u306e\u3067\u3001\u30e2\u30ce\u30af\u30ed\u3068\u30dd\u30ea\u30af\u30ed\u3067\u60a9\u3080\u5fc5\u8981\u306f\u3042\u308a\u307e\u305b\u3093\u3002\r\n\r\n\u3067\u306f\u3001\u4e8c\u6b21\u6297\u4f53\u3067\u306f\u3069\u3046\u3044\u3063\u305f\u70b9\u3092\u30dd\u30a4\u30f3\u30c8\u306b\u9078\u3093\u3067\u3044\u3051\u3070\u3088\u3044\u306e\u304b\u3001\u304a\u8a71\u3092\u7d9a\u3051\u307e\u3057\u3087\u3046\u3002\r\nWhole\u3068F(ab')2\r\n\u4e8c\u6b21\u6297\u4f53\u3092\u63a2\u3057\u3066\u3044\u308b\u3068\u3001\u300cWhole\u300d\u30bf\u30a4\u30d7\u3068\u300cF(ab')2\u300d\u30bf\u30a4\u30d7\u3068\u3044\u3046\u3082\u306e\u3092\u898b\u304b\u3051\u308b\u3053\u3068\u304c\u3042\u308a\u307e\u3059\u3002F(ab')2\u306f\u3001Whole\uff08\u5b8c\u5168\u9577\uff09\u306e\u6297\u4f53\u3092\u30da\u30d7\u30b7\u30f3\u3067\u6d88\u5316\u3057\u305f\u6297\u4f53\u3067\u3059\u3002\u8a00\u3044\u63db\u3048\u308b\u3068\u3001\u6297\u4f53\u306eFc\u90e8\u5206\u3092\u6301\u305f\u306a\u3044\u6297\u4f53\u306e\u3053\u3068\u3067\u3059\u3002\u56f33\u3092\u53c2\u8003\u306b\u3057\u3066\u304f\u3060\u3055\u3044\u3002\r\n\r\n\r\n\r\n\u307e\u305a\u3001\u6297\u4f53\u306e\u6297\u539f\u8a8d\u8b58\u90e8\u4f4d\u306fF(ab')2\u306b\u3042\u308a\u307e\u3059\u306e\u3067\u3001\u6297\u539f\u3078\u306e\u7d50\u5408\u6027\u306fWhole\u3082F(ab')2\u3082\u5909\u308f\u308a\u307e\u305b\u3093\u3002\u4e00\u65b9\u3001F(ab')2\u306b\u306f\u3001Fc\u53d7\u5bb9\u4f53\u306b\u7d50\u5408\u3059\u308bFc\u90e8\u5206\u304c\u306a\u3044\u305f\u3081\u3001\u539f\u7406\u7684\u306b\u306f\u975e\u7279\u7570\u7684\u7d50\u5408\u3092\u6291\u3048\u3089\u308c\u308b\u30e1\u30ea\u30c3\u30c8\u304c\u3042\u308a\u307e\u3059\u304c\u3001\u30a6\u30a7\u30b9\u30bf\u30f3\u30d6\u30ed\u30c3\u30c6\u30a3\u30f3\u30b0\u3067\u306fWhole\u3067\u3082F(ab')2\u3067\u3082\u3042\u307e\u308a\u9055\u3044\u306f\u3042\u308a\u307e\u305b\u3093\u3002\u305d\u306e\u305f\u3081\u3001\u30a6\u30a7\u30b9\u30bf\u30f3\u30d6\u30ed\u30c3\u30c6\u30a3\u30f3\u30b0\u3067\u3042\u308c\u3070\u3069\u3061\u3089\u3092\u4f7f\u3063\u3066\u3082\u554f\u984c\u3042\u308a\u307e\u305b\u3093\u3002\r\n\u306a\u304a\u3001Protein A\u3084Protein G\u306fFc\u306b\u7d50\u5408\u3057\u307e\u3059\u306e\u3067\u3001Protein A\u3084Protein G\u3092\u7528\u3044\u305f\u514d\u75ab\u6c88\u964d\u3067\u306f\u3001F(ab')2\u6297\u4f53\u3092\u4f7f\u308f\u306a\u3044\u3088\u3046\u306b\u6ce8\u610f\u3057\u307e\u3057\u3087\u3046\u3002\r\nCross-Adsorbed\u30bf\u30a4\u30d7\u3068\u306f\uff1f\r\nCross-Adsorbed\u3068\u306f\u3001\u65e5\u672c\u8a9e\u306b\u76f4\u3059\u3068\u300c\u4ea4\u5dee-\u5438\u7740\u3057\u305f\u300d\u6297\u4f53\u3067\u3059\u3002\u3064\u307e\u308a\u3001\u4ea4\u5dee\u53cd\u5fdc\u3092\u8d77\u3053\u3057\u305f\u304f\u306a\u3044\u52d5\u7269\u7a2e\u306e\u6297\u4f53\u3092\u3001\u3042\u3089\u304b\u3058\u3081\u30ab\u30e9\u30e0\u306a\u3069\u306b\u5438\u7740\u3055\u305b\u3001\u53d6\u308a\u9664\u3044\u305f\u6297\u4f53\u3068\u3044\u3046\u610f\u5473\u3067\u3059\u3002\r\n\r\n\u4f8b\u3092\u51fa\u3057\u3066\u3054\u8aac\u660e\u3057\u307e\u3059\u3002Invitrogen\u2122 Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, HRP\u306f\u3001rabbit\u306b\u5bfe\u3059\u308b\u6297\u4f53\u3067\u3059\u304c\u3001human IgG\u3001human serum\u3001mouse IgG\u3001mouse serum\u3001bovine serum\u3092\u30ea\u30ac\u30f3\u30c9\u3068\u3057\u3066\u56fa\u5b9a\u5316\u3057\u305f\u30ec\u30b8\u30f3\u3092\u7528\u3044\u3001\u3053\u308c\u3089\u306b\u53cd\u5fdc\u3059\u308b\u6297\u4f53\u3092\u9664\u53bb\u3057\u305f\u300cCross-Absorbed\u300d\u30bf\u30a4\u30d7\u3067\u3059\u3002\u305d\u306e\u305f\u3081\u3001\u3053\u306e\u6297\u4f53\u306f\u3001human IgG\u3001human serum\u3001mouse IgG\u3001mouse serum\u3001bovine 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Verification\u300d\u3068\u3044\u3046\u30a2\u30a4\u30b3\u30f3\u304c\u3064\u3044\u3066\u3044\u307e\u3059\u3002\u30ce\u30c3\u30af\u30c0\u30a6\u30f3\u30fb\u30ce\u30c3\u30af\u30a2\u30a6\u30c8\u30fb\u4e2d\u548c\u30fbIP\u3068\u8cea\u91cf\u5206\u6790\u6cd5\u306a\u3069\u3001\u898f\u5b9a\u3055\u308c\u305f\u8907\u6570\u306e\u691c\u8a3c\u65b9\u6cd5\u306e\u3046\u3061\u5c11\u306a\u304f\u3068\u30821\u7a2e\u985e\u4ee5\u4e0a\u306e\u691c\u8a3c\u3092\u5b9f\u65bd\u3057\u3066\u3044\u308b\u6297\u4f53\u3067\u3059\u3002\u4f8b\u3048\u3070\u30ce\u30c3\u30af\u30a2\u30a6\u30c8\u691c\u8a3c\u3067\u306f\u3001CRISPR\/Cas9\u3092\u7528\u3044\u3066\u3001\u30bf\u30fc\u30b2\u30c3\u30c8\u30bf\u30f3\u30d1\u30af\u8cea\u306e\u30ce\u30c3\u30af\u30a2\u30a6\u30c8\u682a\u3092\u7528\u3044\u3066\u8a55\u4fa1\u3092\u3057\u3066\u3044\u307e\u3059\u3002\u3082\u3057\u6297\u4f53\u306e\u7279\u7570\u6027\u306b\u4e0d\u5b89\u304c\u3042\u308b\u5834\u5408\u306f\u3001\u3053\u3046\u3044\u3063\u305f\u691c\u8a3c\u6e08\u307f\u306e\u6297\u4f53\u3092\u9078\u629e\u3057\u307e\u3057\u3087\u3046\u3002\r\n\u4e8c\u6b21\u6297\u4f53\u9078\u629e\u6642\u306b\u77e5\u3063\u3066\u304a\u304d\u305f\u3044\u3053\u3068\r\n\u307e\u305a\u3001\u57fa\u672c\u3068\u3057\u3066\u3001\u4e8c\u6b21\u6297\u4f53\u306f\u3001\u4e00\u6b21\u6297\u4f53\u306eHost\uff08\u7531\u6765\u30fb\u514d\u75ab\u52d5\u7269\uff09\u306b\u5fdc\u3058\u3066\u9078\u3073\u307e\u3057\u3087\u3046\u3002\u4f8b\u3048\u3070\u4e00\u6b21\u6297\u4f53\u306eHost\u304cmouse\u3067\u3042\u308c\u3070\u3001\u4e8c\u6b21\u6297\u4f53\u306fAnti-Mouse\u3092\u9078\u629e\u3057\u307e\u3059\u3002\r\n\r\n\u4e8c\u6b21\u6297\u4f53\u3067\u306f\u3001\u4f55\u3089\u304b\u306e\u6a19\u8b58\u304c\u3042\u308b\u3082\u306e\u3092\u7528\u3044\u308b\u3053\u3068\u304c\u4e00\u822c\u7684\u3067\u3059\u3002\u86cd\u5149\u691c\u51fa\u3067\u3042\u308c\u3070\u3001\u86cd\u5149\u6a19\u8b58\u306e\u4e8c\u6b21\u6297\u4f53\u3001\u5316\u5b66\u767a\u5149\u3067\u691c\u51fa\u3059\u308b\u306e\u3067\u3042\u308c\u3070\u3001HRP\u306a\u3069\u306e\u9175\u7d20\u6a19\u8b58\u306e\u3082\u306e\u3092\u7528\u3044\u307e\u3059\u3002\r\n\u307e\u305f\u3001\u66f8\u304b\u308c\u3066\u3044\u306a\u3044\u3053\u3068\u3082\u3042\u308a\u307e\u3059\u304c\u3001\u307b\u3068\u3093\u3069\u306e\u4e8c\u6b21\u6297\u4f53\u306f\u30dd\u30ea\u30af\u30ed\u30fc\u30ca\u30eb\u6297\u4f53\u3067\u3059\u306e\u3067\u3001\u30e2\u30ce\u30af\u30ed\u3068\u30dd\u30ea\u30af\u30ed\u3067\u60a9\u3080\u5fc5\u8981\u306f\u3042\u308a\u307e\u305b\u3093\u3002\r\n\r\n\u3067\u306f\u3001\u4e8c\u6b21\u6297\u4f53\u3067\u306f\u3069\u3046\u3044\u3063\u305f\u70b9\u3092\u30dd\u30a4\u30f3\u30c8\u306b\u9078\u3093\u3067\u3044\u3051\u3070\u3088\u3044\u306e\u304b\u3001\u304a\u8a71\u3092\u7d9a\u3051\u307e\u3057\u3087\u3046\u3002\r\nWhole\u3068F(ab')2\r\n\u4e8c\u6b21\u6297\u4f53\u3092\u63a2\u3057\u3066\u3044\u308b\u3068\u3001\u300cWhole\u300d\u30bf\u30a4\u30d7\u3068\u300cF(ab')2\u300d\u30bf\u30a4\u30d7\u3068\u3044\u3046\u3082\u306e\u3092\u898b\u304b\u3051\u308b\u3053\u3068\u304c\u3042\u308a\u307e\u3059\u3002F(ab')2\u306f\u3001Whole\uff08\u5b8c\u5168\u9577\uff09\u306e\u6297\u4f53\u3092\u30da\u30d7\u30b7\u30f3\u3067\u6d88\u5316\u3057\u305f\u6297\u4f53\u3067\u3059\u3002\u8a00\u3044\u63db\u3048\u308b\u3068\u3001\u6297\u4f53\u306eFc\u90e8\u5206\u3092\u6301\u305f\u306a\u3044\u6297\u4f53\u306e\u3053\u3068\u3067\u3059\u3002\u56f33\u3092\u53c2\u8003\u306b\u3057\u3066\u304f\u3060\u3055\u3044\u3002\r\n\r\n\r\n\r\n\u307e\u305a\u3001\u6297\u4f53\u306e\u6297\u539f\u8a8d\u8b58\u90e8\u4f4d\u306fF(ab')2\u306b\u3042\u308a\u307e\u3059\u306e\u3067\u3001\u6297\u539f\u3078\u306e\u7d50\u5408\u6027\u306fWhole\u3082F(ab')2\u3082\u5909\u308f\u308a\u307e\u305b\u3093\u3002\u4e00\u65b9\u3001F(ab')2\u306b\u306f\u3001Fc\u53d7\u5bb9\u4f53\u306b\u7d50\u5408\u3059\u308bFc\u90e8\u5206\u304c\u306a\u3044\u305f\u3081\u3001\u539f\u7406\u7684\u306b\u306f\u975e\u7279\u7570\u7684\u7d50\u5408\u3092\u6291\u3048\u3089\u308c\u308b\u30e1\u30ea\u30c3\u30c8\u304c\u3042\u308a\u307e\u3059\u304c\u3001\u30a6\u30a7\u30b9\u30bf\u30f3\u30d6\u30ed\u30c3\u30c6\u30a3\u30f3\u30b0\u3067\u306fWhole\u3067\u3082F(ab')2\u3067\u3082\u3042\u307e\u308a\u9055\u3044\u306f\u3042\u308a\u307e\u305b\u3093\u3002\u305d\u306e\u305f\u3081\u3001\u30a6\u30a7\u30b9\u30bf\u30f3\u30d6\u30ed\u30c3\u30c6\u30a3\u30f3\u30b0\u3067\u3042\u308c\u3070\u3069\u3061\u3089\u3092\u4f7f\u3063\u3066\u3082\u554f\u984c\u3042\u308a\u307e\u305b\u3093\u3002\r\n\u306a\u304a\u3001Protein A\u3084Protein G\u306fFc\u306b\u7d50\u5408\u3057\u307e\u3059\u306e\u3067\u3001Protein A\u3084Protein G\u3092\u7528\u3044\u305f\u514d\u75ab\u6c88\u964d\u3067\u306f\u3001F(ab')2\u6297\u4f53\u3092\u4f7f\u308f\u306a\u3044\u3088\u3046\u306b\u6ce8\u610f\u3057\u307e\u3057\u3087\u3046\u3002\r\nCross-Adsorbed\u30bf\u30a4\u30d7\u3068\u306f\uff1f\r\nCross-Adsorbed\u3068\u306f\u3001\u65e5\u672c\u8a9e\u306b\u76f4\u3059\u3068\u300c\u4ea4\u5dee-\u5438\u7740\u3057\u305f\u300d\u6297\u4f53\u3067\u3059\u3002\u3064\u307e\u308a\u3001\u4ea4\u5dee\u53cd\u5fdc\u3092\u8d77\u3053\u3057\u305f\u304f\u306a\u3044\u52d5\u7269\u7a2e\u306e\u6297\u4f53\u3092\u3001\u3042\u3089\u304b\u3058\u3081\u30ab\u30e9\u30e0\u306a\u3069\u306b\u5438\u7740\u3055\u305b\u3001\u53d6\u308a\u9664\u3044\u305f\u6297\u4f53\u3068\u3044\u3046\u610f\u5473\u3067\u3059\u3002\r\n\r\n\u4f8b\u3092\u51fa\u3057\u3066\u3054\u8aac\u660e\u3057\u307e\u3059\u3002Invitrogen\u2122 Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, HRP\u306f\u3001rabbit\u306b\u5bfe\u3059\u308b\u6297\u4f53\u3067\u3059\u304c\u3001human IgG\u3001human serum\u3001mouse IgG\u3001mouse serum\u3001bovine serum\u3092\u30ea\u30ac\u30f3\u30c9\u3068\u3057\u3066\u56fa\u5b9a\u5316\u3057\u305f\u30ec\u30b8\u30f3\u3092\u7528\u3044\u3001\u3053\u308c\u3089\u306b\u53cd\u5fdc\u3059\u308b\u6297\u4f53\u3092\u9664\u53bb\u3057\u305f\u300cCross-Absorbed\u300d\u30bf\u30a4\u30d7\u3067\u3059\u3002\u305d\u306e\u305f\u3081\u3001\u3053\u306e\u6297\u4f53\u306f\u3001human IgG\u3001human serum\u3001mouse IgG\u3001mouse serum\u3001bovine 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>お問い合わせ</span></a></li> </ul></nav></div></section> </div><div class="breadcrumb"><span class="breadcrumb-link-wrap"><a href="https://www.thermofisher.com/blog/?">Accelerating Science</a></span><span class="breadcrumb-link-wrap"><a class="breadcrumb-link" href="https://www.thermofisher.com/blog/learning-at-the-bench/"><span class="breadcrumb-link-text-wrap">Learning at the Bench</span></a><meta ></span> <span aria-label="breadcrumb separator">/</span> <span class="breadcrumb-link-wrap"><a class="breadcrumb-link" href="https://www.thermofisher.com/blog/learning-at-the-bench/mol-bio/"><span class="breadcrumb-link-text-wrap">分子生物学実験関連</span></a><meta ></span> <span aria-label="breadcrumb separator">/</span> <span class="breadcrumb-current-title">一次抗体と二次抗体を選ぶ上で知っておきたいこと：ウェスタンブロッティングの選び方　第6回</span></div></div></header><div class="site-inner"><div class="content-sidebar-wrap"><main class="content"><article class="post-15396 post type-post status-publish format-standard has-post-thumbnail category-mol-bio tag-western-blotting entry" aria-label="一次抗体と二次抗体を選ぶ上で知っておきたいこと：ウェスタンブロッティングの選び方　第6回"><header class="entry-header"><h1 class="entry-title">一次抗体と二次抗体を選ぶ上で知っておきたいこと：ウェスタンブロッティングの選び方　第6回</h1> <span class="post-info-author"> 作成者 <a href="https://www.thermofisher.com/blog/author/lgbadmin/" title="LatB Staff の投稿" rel="author">LatB Staff</a> </span> <span class="post-info-date"> 09.26.2023 </span> </header><div class="entry-content"><p><img decoding="async" class="aligncenter size-full wp-image-13122" src="https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2022/12/western.jpg" alt="westernblotting" width="761" height="476" srcset="https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2022/12/western.jpg 761w, https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2022/12/western-300x188.jpg 300w, https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2022/12/western-320x200.jpg 320w" sizes="(max-width: 761px) 100vw, 761px" /><br /> ウェスタンブロッティングの「選び方」について連載している本シリーズ、<a href="https://www.thermofisher.com/blog/learning-at-the-bench/western-blot-protein-ts-5/?CID=bid_pca_wwg_r04_jp_cp0000_pjt0000_bid00000_0so_blg_op_awa_oc_western_blot_protein_ts_6_Social_LAB" target="_blank" rel="noopener">前回</a>はブロッキングについてご紹介しました。ブロッキングの次のステップは抗原抗体反応、つまり一次抗体と二次抗体が必要になります。<br /> ここでは、抗体を選ぶときに知っておきたいポイントについて、ご紹介します。</p> <div id="toc_container" class="no_bullets"><p class="toc_title">▼もくじ</p><ul class="toc_list"><li><a href="#i">一次抗体選択時に知っておきたいこと</a><ul><li><a href="#i-2">モノクロとポリクロ</a></li><li><a href="#i-3">アプリケーション確認は必須</a></li><li><a href="#i-4">特異性検証済みの一次抗体も</a></li></ul></li><li><a href="#i-5">二次抗体選択時に知っておきたいこと</a><ul><li><a href="#WholeFab82172">WholeとF(ab’)2</a></li><li><a href="#Cross-Adsorbed">Cross-Adsorbedタイプとは？</a></li></ul></li><li><a href="#i-6">抗体希釈液と濃度</a></li><li><a href="#i-7">一次抗体の探し方</a></li><li><a href="#i-8">まとめ</a><ul><li><a href="#i-9">【無料ダウンロード】タンパク質解析ワークフローハンドブック</a></li></ul></li></ul></div> <h2><span id="i">一次抗体選択時に知っておきたいこと</span></h2> <p>ウェスタンブロッティングでは、ブロッキングを終えた後、一次抗体でターゲットタンパク質を特異的に認識させ、酵素や蛍光標識されている二次抗体を一次抗体に反応させます。その後、蛍光標識二次抗体の場合は検出ステップに移り、酵素標識二次抗体の場合は、化学発光や発色などの基質を酵素に反応させて、検出を行います。ここまでの流れを終えたメンブレンの状態を、図1に示しました。</p> <div id="attachment_15401" style="width: 628px" class="wp-caption aligncenter"><img decoding="async" aria-describedby="caption-attachment-15401" class="size-full wp-image-15401" src="https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-01.jpg" alt="ウェスタンブロッティング二次抗体反応終了後のメンブレンの状態" width="618" height="220" srcset="https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-01.jpg 618w, https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-01-300x107.jpg 300w" sizes="(max-width: 618px) 100vw, 618px" /><p id="caption-attachment-15401" class="wp-caption-text">図1：ウェスタンブロッティング二次抗体反応終了後のメンブレンの状態</p></div> <p>本ブログでクローズアップするのは、目的タンパク質に結合する一次抗体と、酵素や蛍光色素などが標識された二次抗体についてです。</p> <p>抗体を自作するのは大変なので、市販品を用意されている人がほとんどでしょう。しかしメーカーのサイトで一次抗体や二次抗体を検索してみると、複数ヒットしてしまい、どれが良いのかわからないことも多いと思います。</p> <p>そういうときのために、本ブログでは、「抗体選択時に知っておきたいこと」をまとめました。</p> <h3><span id="i-2"><u>モノクロとポリクロ</u></span></h3> <p>一次抗体には、モノクロ―ナル抗体とポリクローナル抗体があります。モノクロ―ナル抗体は、抗原を注射した動物から、抗体を産生するB細胞を単離して、ミエローマという培養細胞と融合させて抗体を作製したもので、単一（モノ）のエピトープ部位を認識する抗体です。ポリクローナル抗体は、動物に抗原を複数回注射した後、血液を採取し、その血清から抗体を精製したもので、それぞれ異なるエピトープ部位を認識する複数（ポリ）の抗体の混合物です。</p> <div id="attachment_15399" style="width: 507px" class="wp-caption aligncenter"><img loading="lazy" decoding="async" aria-describedby="caption-attachment-15399" class="size-full wp-image-15399" src="https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-02.jpg" alt="モノクロ―ナル抗体とポリクローナル抗体" width="497" height="204" srcset="https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-02.jpg 497w, https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-02-300x123.jpg 300w" sizes="(max-width: 497px) 100vw, 497px" /><p id="caption-attachment-15399" class="wp-caption-text">図2：モノクローナル抗体とポリクローナル抗体</p></div> <p>では、これらの選択によって、ウェスタンブロッティングではどういう違いがもたらされるのでしょうか。両者を「非特異反応」と「検出感度」において比較すると、次のようになります。</p> <p><b>◆ウェスタンブロッティングにおける非特異的反応の起こりにくさ◆</b><br /> <b>モノクロ―ナル抗体　＞　ポリクローナル抗体</b></p> <p><b>◆ウェスタンブロッティングにおける一般的な検出感度◆</b><br /> <b>ポリクローナル抗体　＞　モノクロ―ナル抗体</b></p> <p>非特異反応と検出感度、どちらもウェスタンブロッティングでは重要な要素ですが、モノクロ―ナル抗体とポリクローナル抗体では、相反する性質を持っています。</p> <p>もう少し詳しく解説しましょう。<br /> ポリクローナル抗体の交差性がモノクロ―ナル抗体より大きいのは、複数の抗体のプールですので、一般的な傾向として、相対的に非特異反応を持つ抗体を含むリスクが高いためです。しかし、1つの抗原に複数の抗体が結合できるので、二次抗体も複数結合できます。それゆえに、ポリクローナル抗体では、非特異的反応のリスクは比較的高くなるものの、検出感度は高くなりやすい傾向にあります。モノクロ―ナル抗体はこれの逆で、非特異的反応は低いですが、検出感度はポリクローナル抗体と比べると低くなることがあります。これらの特性を考えて、どちらを選ぶのかを決めましょう。</p> <h3><span id="i-3"><u>アプリケーション確認は必須</u></span></h3> <p>多くの抗体メーカーでは、その一次抗体がどのアプリケーションに適しているか書かれています。この内容は抗体選択時に必ず確認しましょう。<br /> アプリケーション確認が必須なのは、アプリケーションによって抗原の状態が異なるためです。例えばELISAや免疫染色では、タンパク質の形が保たれた抗原を、抗体が認識します。一方、ウェスタンブロッティングでは、SDSによって変性し、DTTによって還元処理された抗原を、抗体が認識しなくてはなりません。そのため、例えばELISAで確認済みの抗体では、ウェスタンブロッティングで抗原を検出できないことがあります。一次抗体を選ぶときは、アプリケーション欄をしっかり確認しておきましょう。</p> <h3><span id="i-4"><u>特異性検証済みの一次抗体も</u></span></h3> <p>ウェスタンブロッティングで無事に検出できたけれど、果たしてこのバンドは本当に目的タンパク質だろうか？そんな疑問と向き合った経験がある方もいるかもしれません。抗体の特異性や性能について実際に検証された抗体を使いたいという研究者は多くいらっしゃいます。こういった背景から、抗体が正しくターゲットと結合したことを証明した「検証済みの抗体」というものが存在します。</p> <p>メーカーによって呼び名は異なりますが、たとえば、サーモフィッシャーサイエンティフィックでは、「<a href="https://www.thermofisher.com/jp/ja/home/life-science/antibodies/invitrogen-antibody-validation.html?CID=bid_pca_wwg_r04_jp_cp0000_pjt0000_bid00000_0so_blg_op_awa_oc_western_blot_protein_ts_6_Social_LAB" target="_blank" rel="noopener">Advanced Verification</a>」というアイコンがついています。ノックダウン・ノックアウト・中和・IPと質量分析法など、規定された複数の検証方法のうち少なくとも1種類以上の検証を実施している抗体です。例えばノックアウト検証では、CRISPR/Cas9を用いて、ターゲットタンパク質のノックアウト株を用いて評価をしています。もし抗体の特異性に不安がある場合は、こういった検証済みの抗体を選択しましょう。</p> <h2><span id="i-5">二次抗体選択時に知っておきたいこと</span></h2> <p>まず、基本として、二次抗体は、一次抗体のHost（由来・免疫動物）に応じて選びましょう。例えば一次抗体のHostがmouseであれば、二次抗体はAnti-Mouseを選択します。</p> <p>二次抗体では、何らかの標識があるものを用いることが一般的です。蛍光検出であれば、蛍光標識の二次抗体、化学発光で検出するのであれば、HRPなどの酵素標識のものを用います。<br /> また、書かれていないこともありますが、ほとんどの二次抗体はポリクローナル抗体ですので、モノクロとポリクロで悩む必要はありません。</p> <p>では、二次抗体ではどういった点をポイントに選んでいけばよいのか、お話を続けましょう。</p> <h3><span id="WholeFab82172"><u>WholeとF(ab’)2</u></span></h3> <p>二次抗体を探していると、「Whole」タイプと「F(ab’)2」タイプというものを見かけることがあります。F(ab’)2は、Whole（完全長）の抗体をペプシンで消化した抗体です。言い換えると、抗体のFc部分を持たない抗体のことです。図3を参考にしてください。</p> <div id="attachment_15400" style="width: 412px" class="wp-caption aligncenter"><img loading="lazy" decoding="async" aria-describedby="caption-attachment-15400" class="size-full wp-image-15400" src="https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-03.png" alt="抗体断片の名称" width="402" height="125" srcset="https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-03.png 402w, https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-03-300x93.png 300w" sizes="(max-width: 402px) 100vw, 402px" /><p id="caption-attachment-15400" class="wp-caption-text">図3：抗体断片の名称</p></div> <p>まず、抗体の抗原認識部位はF(ab’)2にありますので、抗原への結合性はWholeもF(ab’)2も変わりません。一方、F(ab’)2には、Fc受容体に結合するFc部分がないため、原理的には非特異的結合を抑えられるメリットがありますが、ウェスタンブロッティングではWholeでもF(ab’)2でもあまり違いはありません。そのため、ウェスタンブロッティングであればどちらを使っても問題ありません。<br /> なお、Protein AやProtein GはFcに結合しますので、Protein AやProtein Gを用いた免疫沈降では、F(ab’)2抗体を使わないように注意しましょう。</p> <h3><span id="Cross-Adsorbed"><u>Cross-Adsorbedタイプとは？</u></span></h3> <p>Cross-Adsorbedとは、日本語に直すと「交差-吸着した」抗体です。つまり、交差反応を起こしたくない動物種の抗体を、あらかじめカラムなどに吸着させ、取り除いた抗体という意味です。</p> <p>例を出してご説明します。Invitrogen™ <a href="https://www.thermofisher.com/antibody/product/Goat-anti-Rabbit-IgG-H-L-Cross-Adsorbed-Secondary-Antibody-Polyclonal/G-21234?CID=bid_pca_wwg_r04_jp_cp0000_pjt0000_bid00000_0so_blg_op_awa_oc_western_blot_protein_ts_6_Social_LAB" target="_blank" rel="noopener">Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, HRP</a>は、rabbitに対する抗体ですが、human IgG、human serum、mouse IgG、mouse serum、bovine serumをリガンドとして固定化したレジンを用い、これらに反応する抗体を除去した「Cross-Absorbed」タイプです。そのため、この抗体は、human IgG、human serum、mouse IgG、mouse serum、bovine serumへの交差性を持たない抗体と言えます。</p> <p>Cross-Adsorbedタイプの抗体として適切なものを選べば、サンプル中の内在性IgGへの反応を防ぐことができます。また、例えば1枚のメンブレンで複数のバンドを多重検出する際、異なる動物種由来の一次抗体を用いるケースでも、安心して使用できます（蛍光ウェスタンブロッティング多重染色については<a href="https://www.thermofisher.com/jp/ja/home/life-science/protein-biology/protein-assays-analysis/western-blotting/detect-proteins-western-blot/fluorescent-western-blot-detection.html?CID=bid_pca_wwg_r04_jp_cp0000_pjt0000_bid00000_0so_blg_op_awa_oc_western_blot_protein_ts_6_Social_LAB" target="_blank" rel="noopener">こちら</a>）。</p> <p>また、二次抗体も、同様にアプリケーションとしてウェスタンブロッティングで確認済みのものを使用しましょう。</p> <h2><span id="i-6">抗体希釈液と濃度</span></h2> <p>さて、抗体が決まったら、抗体を希釈する液を用意しましょう。表1に示した3種類の溶液が、一次抗体や二次抗体の希釈液としてよく使われています。</p> <table> <caption>表1. 抗体の希釈液</caption> <tbody> <tr> <td width="132">抗体希釈液</td> <td width="454">内容</td> </tr> <tr> <td width="132">TBS-T</td> <td width="454">TBSに0.1%のTween 20を加えたもの</td> </tr> <tr> <td width="132">PBS-T</td> <td width="454">PBSに0.1%のTween 20を加えたもの<br /> ※リン酸化タンパク質の検出には使用できない</td> </tr> <tr> <td width="132">ブロッキング試薬</td> <td width="454">抗原抗体反応前のブロッキングに用いた試薬</td> </tr> </tbody> </table> <p>TBS-TまたはPBS-TにTween™ 20を加えることで、メンブレン上の抗原以外の部分に抗体が結合するのを抑える、つまり非特異的反応を抑えるメリットがあります。<br /> また、ブロッキング試薬で抗体を希釈することで、抗体反応を行いながらブロッキングも実施し、こちらも非特異的反応を軽減させる効果があります（抗体希釈液にブロッキング試薬を使うときも、転写後のブロッキングステップは必須です）。しかし、抗体希釈液へブロッキング試薬や界面活性剤を添加しすぎると、抗体の抗原への結合も抑えられてしまい、ターゲットのシグナル低減に結びつくこともありますので、万能な方法ではないことに注意しましょう。まずは上述のいずれかで実施し、結果を見てみることをおすすめします。</p> <p>次に、抗体の希釈濃度です。</p> <p>たいていの一次抗体や二次抗体には、ウェスタンブロッティングの希釈倍率の目安が書かれています。しかし、<b><u>抗体推奨の希釈倍率ではなく、検出試薬推奨の希釈倍率に沿うことをおすすめします。</u></b><br /> これは、抗体の推奨希釈倍率は、検出試薬の感度を考慮していないことが多いためです。ウェスタンブロッティングでは、検出感度は検出試薬によって大きく変わります。検出感度が高いのに抗体濃度が高すぎるとバックグラウンドが真っ黒になったり、逆に検出感度が低いのに抗体濃度が低すぎるとバンドが検出されなかったりします。</p> <p>例として、実際の製品の情報を見ながら解説しましょう。下表は、二次抗体であるInvitrogen™ <a href="https://www.thermofisher.com/antibody/product/Goat-anti-Rabbit-IgG-H-L-Secondary-Antibody-Polyclonal/65-6120?CID=bid_pca_wwg_r04_jp_cp0000_pjt0000_bid00000_0so_blg_op_awa_oc_western_blot_protein_ts_6_Social_LAB" target="_blank" rel="noopener">Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP</a>の推奨希釈倍率です。<img loading="lazy" decoding="async" class="aligncenter size-full wp-image-15403" src="https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-04.jpg" alt="" width="789" height="121" srcset="https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-04.jpg 789w, https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-04-300x46.jpg 300w, https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-04-768x118.jpg 768w" sizes="(max-width: 789px) 100vw, 789px" /><br /> つまり、この抗体の情報として、ウェスタンブロッティングでは3,000倍から10,000倍希釈してくださいと書かれています。</p> <p>一方、検出試薬に書かれている二次抗体の希釈倍率を見てみましょう。</p> <table> <caption>表2. 検出試薬ごとの抗体希釈倍率</caption> <tbody> <tr> <td width="104">通称</td> <td width="216">製品リンク</td> <td width="105">一次抗体</td> <td width="170">二次抗体</td> </tr> <tr> <td width="104">West Atto</td> <td width="216">Thermo Scientific™<br /> <a href="https://www.thermofisher.com/order/catalog/product/A38555?CID=bid_pca_wwg_r04_jp_cp0000_pjt0000_bid00000_0so_blg_op_awa_oc_western_blot_protein_ts_6_Social_LAB" target="_blank" rel="noopener">SuperSignal™ West Atto Ultimate Sensitivity Substrate</a></td> <td width="105">1:5,000</td> <td width="170">1:100,000 ～ 1:250,000</td> </tr> <tr> <td width="104">West Femto</td> <td width="216">Thermo Scientific™<br /> <a href="https://www.thermofisher.com/order/catalog/product/34095?CID=bid_pca_wwg_r04_jp_cp0000_pjt0000_bid00000_0so_blg_op_awa_oc_western_blot_protein_ts_6_Social_LAB" target="_blank" rel="noopener">SuperSignal™ West Femto Maximum Sensitivity Substrate</a></td> <td width="105">1:5,000</td> <td width="170">1:100,000 ～ 1:500,000</td> </tr> <tr> <td width="104">West Dura</td> <td width="216">Thermo Scientific™<br /> <a href="https://www.thermofisher.com/order/catalog/product/34075?CID=bid_pca_wwg_r04_jp_cp0000_pjt0000_bid00000_0so_blg_op_awa_oc_western_blot_protein_ts_6_Social_LAB" target="_blank" rel="noopener">SuperSignal™ West Dura Extended Duration Substrate</a></td> <td width="105">1:5,000</td> <td width="170">1:50,000 ～ 1:250,000</td> </tr> <tr> <td width="104">West Pico PLUS</td> <td width="216">Thermo Scientific™<br /> <a href="https://www.thermofisher.com/order/catalog/product/34577?CID=bid_pca_wwg_r04_jp_cp0000_pjt0000_bid00000_0so_blg_op_awa_oc_western_blot_protein_ts_6_Social_LAB" target="_blank" rel="noopener">SuperSignal™ West Pico PLUS Chemiluminescent Substrate</a></td> <td width="105">1:1,000</td> <td width="170">1:20,000 ～ 1:100,000</td> </tr> <tr> <td width="104">Pierce ECL Plus</td> <td width="216">Thermo Scientific™<br /> <a href="https://www.thermofisher.com/order/catalog/product/32132?CID=bid_pca_wwg_r04_jp_cp0000_pjt0000_bid00000_0so_blg_op_awa_oc_western_blot_protein_ts_6_Social_LAB" target="_blank" rel="noopener">Pierce™ ECL Plus Western Blotting Substrate</a></td> <td width="105">1:1,000</td> <td width="170">1:25,000 ～ 1:200,000</td> </tr> <tr> <td width="104">Pierce ECL</td> <td width="216">Thermo Scientific™<br /> <a href="https://www.thermofisher.com/order/catalog/product/32209?CID=bid_pca_wwg_r04_jp_cp0000_pjt0000_bid00000_0so_blg_op_awa_oc_western_blot_protein_ts_6_Social_LAB" target="_blank" rel="noopener">Pierce™ ECL Western Blotting Substrate</a></td> <td width="105">1:1,000</td> <td width="170">1:1,000 ～ 1:15,000</td> </tr> </tbody> </table> <p>表2を見ると、検出試薬によって抗体の希釈倍率が大きく異なることがわかります。<a href="https://www.thermofisher.com/antibody/product/Goat-anti-Rabbit-IgG-H-L-Secondary-Antibody-Polyclonal/65-6120?CID=bid_pca_wwg_r04_jp_cp0000_pjt0000_bid00000_0so_blg_op_awa_oc_western_blot_protein_ts_6_Social_LAB" target="_blank" rel="noopener">Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP</a>を使用するとき、この抗体に書かれている希釈倍率3,000～10,000倍を信じて、例えばWest Attoで検出をするとします。しかし、West Attoの推奨二次抗体希釈倍率は100,000～250,000倍なので、3,000倍希釈では二次抗体が濃すぎて、バックグラウンドがかなり高くなると予想されます。このように、トラブルの原因になることがありますので、抗体は、検出試薬の推奨条件に従って希釈してください。</p> <p>また、抗体の希釈倍率は、一般的に、抗体の原液が1 mg/mＬであることを前提に書かれています。もし購入した抗体の原液濃度が1 mg/mＬではない場合、希釈倍率も濃度に応じて計算してください。</p> <h2><span id="i-7">一次抗体の探し方</span></h2> <p>では、実際に抗体をWebから探してみましょう。サーモフィッシャーサイエンティフィックのサイトの左上にある「Search All」のプルダウンをクリックし、一次抗体を選択します。<img loading="lazy" decoding="async" class="aligncenter size-full wp-image-15406" src="https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-05.jpg" alt="" width="906" height="272" srcset="https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-05.jpg 906w, https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-05-300x90.jpg 300w, https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-05-768x231.jpg 768w" sizes="(max-width: 906px) 100vw, 906px" /><br /> すると、下図のように検索バーが変わります。<img loading="lazy" decoding="async" class="aligncenter size-full wp-image-15407" src="https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-06.jpg" alt="" width="946" height="85" srcset="https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-06.jpg 946w, https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-06-300x27.jpg 300w, https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-06-768x69.jpg 768w" sizes="(max-width: 946px) 100vw, 946px" /><br /> 「Target of interest(Target, gene, symbol, antigen)」にタンパク質名を入力、「Application」のプルダウンからアプリケーションを選択、「Target Species」のプルダウンからターゲットの動物種を選択し、右端の検索ボタンを押してください。</p> <p>検索結果が表示されます。<br /> さらに絞り込みを行いたい場合は、下図赤枠から、検索結果をフィルタリングできます。<img loading="lazy" decoding="async" class="aligncenter size-full wp-image-15404" src="https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-07.jpg" alt="" width="1713" height="287" srcset="https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-07.jpg 1713w, https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-07-300x50.jpg 300w, https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-07-1024x172.jpg 1024w, https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-07-768x129.jpg 768w, https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-07-1536x257.jpg 1536w" sizes="(max-width: 1713px) 100vw, 1713px" /><br /> 例えば、ポリクローナル抗体に絞り込みたい場合は、「Clonality」から「Polyclonal」を選択してください。<img loading="lazy" decoding="async" class="aligncenter wp-image-15405" src="https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-08.jpg" alt="" width="280" height="235" srcset="https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-08.jpg 346w, https://www.thermofisher.com/blog/learning-at-the-bench/wp-content/uploads/sites/13/2023/09/western-blot-protein-ts-6-08-300x252.jpg 300w" sizes="(max-width: 280px) 100vw, 280px" /><br /> 二次抗体も、同様に「Search All」のプルダウンから二次抗体を選んで、検索結果をフィルタリングして絞り込みが可能です。</p> <h2><span id="i-8">まとめ</span></h2> <p>ウェスタンブロッティングにおける抗体の選び方のポイントをまとめました。</p> <ul> <li>一次抗体では、モノクロ―ナル抗体では非特異的反応が起こりにくく、一般的な検出感度はポリクローナル抗体の方が高くなりやすい</li> <li>抗体は、ウェスタンブロッティングで確認済みのものを用いる</li> <li>二次抗体では、F(ab’)2抗体を用いることで非特異反応を抑えたり、Cross-Absorbedタイプでほかの生物種に交差性のない抗体を手に入れたりすることができる</li> <li>抗体は、TBS-TやPBS-T、ブロッキング試薬などで希釈し、その希釈倍率は検出試薬の推奨に沿う</li> </ul> <p><a href="https://www.thermofisher.com/blog/learning-at-the-bench/western-blot-protein-ts-7/?CID=bid_pca_wwg_r04_jp_cp0000_pjt0000_bid00000_0so_blg_op_awa_oc_western_blot_protein_ts_6_Social_LAB" rel="noopener" target="_blank">次回</a>は、検出試薬選択のポイントについてご紹介します。</p> <p>ウェスタンブロッティングの選び方シリーズはこちら：<br /> <a href="https://www.thermofisher.com/blog/learning-at-the-bench/western-blot-protein-ts-1/?CID=bid_pca_wwg_r04_jp_cp0000_pjt0000_bid00000_0so_blg_op_awa_oc_western_blot_protein_ts_6_Social_LAB" target="_blank" rel="noopener">ウェスタンブロッティングの製品選びにもう困らない！第1回：タンパク質の抽出方法</a><br /> <a href="https://www.thermofisher.com/blog/learning-at-the-bench/western-blot-protein-ts-2/?CID=bid_pca_wwg_r04_jp_cp0000_pjt0000_bid00000_0so_blg_op_awa_oc_western_blot_protein_ts_6_Social_LAB" target="_blank" rel="noopener">ウェスタンブロッティングの製品選びにもう困らない！第2回：総タンパク質定量キット</a><br /> <a href="https://www.thermofisher.com/blog/learning-at-the-bench/western-blot-protein-ts-3/?CID=bid_pca_wwg_r04_jp_cp0000_pjt0000_bid00000_0so_blg_op_awa_oc_western_blot_protein_ts_6_Social_LAB" target="_blank" rel="noopener">SDS-PAGEの選び方：ウェスタンブロッティングの製品選びにもう困らない！第3回</a><br /> <a href="https://www.thermofisher.com/blog/learning-at-the-bench/western-blot-protein-ts-4/?CID=bid_pca_wwg_r04_jp_cp0000_pjt0000_bid00000_0so_blg_op_awa_oc_western_blot_protein_ts_6_Social_LAB" target="_blank" rel="noopener">転写条件の選び方：ウェスタンブロッティングの製品選びにもう困らない！第4回</a><br /> <a href="https://www.thermofisher.com/blog/learning-at-the-bench/western-blot-protein-ts-5/?CID=bid_pca_wwg_r04_jp_cp0000_pjt0000_bid00000_0so_blg_op_awa_oc_western_blot_protein_ts_6_Social_LAB" target="_blank" rel="noopener">ブロッキング試薬のポイント：ウェスタンブロッティングの製品選びにもう困らない！第5回</a><br /> 抗体を選ぶときに知っておきたいこと：ウェスタンブロッティングの製品選び第6回<br /> <a href="https://www.thermofisher.com/blog/learning-at-the-bench/western-blot-protein-ts-7/?CID=bid_pca_wwg_r04_jp_cp0000_pjt0000_bid00000_0so_blg_op_awa_oc_western_blot_protein_ts_6_Social_LAB" rel="noopener" target="_blank">化学発光とマルチプレックス検出の選択ポイント：ウェスタンブロッティングの選び方　第7回</a></p> <h3><span id="i-9">【無料ダウンロード】タンパク質解析ワークフローハンドブック</span></h3> 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一次抗体と二次抗体を選ぶ上で知っておきたいこと：ウェスタンブロッティングの選び方 第6回 - Learning at the Bench

一次抗体と二次抗体を選ぶ上で知っておきたいこと：ウェスタンブロッティングの選び方　第6回 - Learning at the Bench