{"id":180,"date":"2023-03-29T02:15:29","date_gmt":"2023-03-28T18:15:29","guid":{"rendered":"http:\/\/39.105.184.3\/?p=180"},"modified":"2025-12-21T01:06:30","modified_gmt":"2025-12-20T17:06:30","slug":"%e8%bf%9e%e7%bb%ad%e5%81%9a%e4%ba%86%e5%8d%81%e5%87%a0%e4%b8%aawestern-blot%e7%9a%84%e7%bb%8f%e9%aa%8c%e8%ae%b0%e5%bd%95","status":"publish","type":"post","link":"https:\/\/fanyiming.life\/?p=180","title":{"rendered":"\u8fde\u7eed\u505a\u4e86\u5341\u51e0\u4e2aWestern blot\u7684\u7ecf\u9a8c\u8bb0\u5f55"},"content":{"rendered":"\n<h2 class=\"wp-block-heading\">\u5173\u4e8e\u5c0f\u5206\u5b50\u86cb\u767d\u5728 12% TG \u80f6\u4e2d\u7684\u5206\u79bb<\/h2>\n\n\n\n<p><strong>\u5b9e\u9a8c\u80cc\u666f<\/strong>\uff082025.8.29\uff09<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>\u6837\u54c1\uff1aK562 \u7ec6\u80de\u88c2\u89e3\u6db2\uff0c\u76ee\u6807\u68c0\u6d4b \u03b3-H2AX (~14 kDa)\u3002<\/li>\n\n\n\n<li>\u7535\u6cf3\u6761\u4ef6\uff1a12% Tris-Glycine SDS-PAGE\uff0c\u5e38\u89c4\u7535\u538b\u3002<\/li>\n\n\n\n<li>\u89c2\u5bdf\uff1a\u5206\u5b50\u91cf\u6807\u8bb0\uff08marker\uff09\u4e2d\uff0c\u5927\u5206\u5b50\uff08&gt;40 kDa\uff09\u5728\u5206\u79bb\u80f6\u4e0a\u5f88\u5feb\u5c31\u5c55\u5f00\u5206\u79bb\uff0c\u800c 10 kDa \u4e0e 15 kDa \u7684\u6761\u5e26\u4e00\u76f4\u5230\u63a5\u8fd1\u80f6\u5e95\u65f6\u624d\u5f7c\u6b64\u62c9\u5f00\u3002<\/li>\n<\/ul>\n\n\n\n<p><strong>\u73b0\u8c61\u603b\u7ed3<\/strong><\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>\u5728 12% TG \u80f6\u4f53\u7cfb\u4e2d\uff0c\u5c0f\u5206\u5b50\uff0810\u201315 kDa\uff09\u8fc1\u79fb\u51e0\u4e4e\u7d27\u8ddf\u6eb4\u915a\u84dd\u524d\u6cbf\uff0c\u5f88\u665a\u624d\u771f\u6b63\u88ab\u201c\u7b5b\u5f00\u201d\uff1b<\/li>\n\n\n\n<li>\u5bf9\u6bd4\u4e4b\u4e0b\uff0c\u4e2d\u5927\u5206\u5b50\u86cb\u767d\uff0840\u201370 kDa\uff09\u5728\u5206\u79bb\u80f6\u4e0a\u6bb5\u5c31\u80fd\u83b7\u5f97\u6e05\u6670\u5206\u79bb\u3002<\/li>\n<\/ul>\n\n\n\n<p><strong>\u673a\u5236\u7406\u89e3<\/strong><\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>Tris-Glycine \u4f53\u7cfb\u4e2d\uff0c\u79bb\u5b50\u524d\u6cbf\u6548\u5e94\u51b3\u5b9a\u4e86\u86cb\u767d\u6700\u521d\u7684\u201c\u5806\u79ef\u2014\u89e3\u5806\u79ef\u201d\u8fc7\u7a0b\u3002<\/li>\n\n\n\n<li>\u5bf9 &lt;20 kDa \u7684\u86cb\u767d\uff0c\u5206\u79bb\u80f6\u5b54\u5f84\u4ecd\u7136\u504f\u5927 \u2192 \u5728\u5927\u90e8\u5206\u8fc1\u79fb\u8fc7\u7a0b\u4e2d\u7b5b\u5206\u6548\u5e94\u4e0d\u5f3a\uff1b<\/li>\n\n\n\n<li>\u540c\u65f6 Gly \u5728 pH 8.8 \u4e0b\u8fc1\u79fb\u901f\u5ea6\u52a0\u5feb\uff0c\u5c0f\u86cb\u767d\u8fc1\u79fb\u7387\u63a5\u8fd1\u79bb\u5b50\u524d\u6cbf \u2192 \u957f\u65f6\u95f4\u51e0\u4e4e\u4e0d\u5206\u5f00\uff0c\u76f4\u5230\u8dd1\u5230\u80f6\u5e95\u624d\u5c55\u73b0\u5dee\u5f02\u3002<\/li>\n<\/ul>\n\n\n\n<p><strong>\u7ecf\u9a8c\u7ed3\u8bba<\/strong><\/p>\n\n\n\n<p>\u540c\u65f6\u5e94\u63a7\u5236\u7535\u538b\u4e0e\u6e29\u5ea6\uff0c\u907f\u514d\u524d\u6cbf\u8fc7\u5feb\u63a8\u8fdb\u5bfc\u81f4\u5c0f\u5206\u5b50\u201c\u8ddf\u7740\u6eb4\u915a\u84dd\u4e00\u8d77\u8dd1\u201d\u3002<\/p>\n\n\n\n<p><strong>12% TG \u80f6\u5e76\u4e0d\u9002\u5408\u5206\u8fa8 10\u201315 kDa \u7684\u86cb\u767d<\/strong>\uff0c\u8fd9\u4f1a\u5bfc\u81f4 \u03b3-H2AX \u7b49\u5c0f\u5206\u5b50\u76ee\u6807\u5728\u9760\u8fd1\u80f6\u5e95\u624d\u5206\u79bb\uff0c\u5206\u8fa8\u7387\u6709\u9650\u3002<\/p>\n\n\n\n<p>\u82e5\u76ee\u6807\u86cb\u767d &lt;20 kDa\uff0c\u63a8\u8350\u4f7f\u7528\uff1a<\/p>\n\n\n\n<p><strong>Tris-Tricine SDS-PAGE\uff0816\u201316.5%\uff09<\/strong> \u6216<\/p>\n\n\n\n<p><strong>Bis-Tris + MES \u4f53\u7cfb<\/strong> \u6216<\/p>\n\n\n\n<p><strong>\u9ad8\u6d53\u5ea6\/\u68af\u5ea6 TG \u80f6\uff0815\u201316.5% \u6216 4\u201320%\uff09<\/strong>\u3002<\/p>\n\n\n\n<h2 class=\"wp-block-heading\">\u53ef\u91cd\u590d\u6027\u95ee\u9898<\/h2>\n\n\n\n<p>\u6c38\u8fdc\u4e0d\u8981\u76f8\u4fe1\u5355\u6b21WB\u7684\u5b9e\u9a8c\u7ed3\u679c\uff0c\u4e5f\u4e0d\u8981\u5bf9\u7740\u7ed3\u679c\u8fc7\u5ea6\u5206\u6790\uff0c\u5728\u91cd\u590d\u4e09\u6b21\u4ee5\u4e0a\u4e4b\u524d\u3002\u8fd9\u6b21\u6b63\u597d\u6709\u4e2aWT\u548cKO\u6837\u54c1\u641e\u53cd\u4e86\u7684\u4e8b\u6545\uff0c\u6240\u4ee5\u7528\u5b8c\u5168\u4e00\u6837\u7684\u6837\u54c1\u53c8\u8dd1\u4e86\u4e00\u6b21\uff0c\u7ed3\u679c\u628a\u4e24\u6b21\u7684\u7ed3\u679c\u653e\u5728\u4e00\u8d77\u5bf9\u6bd4\uff0c\u53d1\u73b0\u5bf9\u7167\u4e4b\u95f4\u7684\u4e0d\u5c11\u6761\u5e26\u76f8\u5bf9\u4eae\u5ea6\u5728\u4e24\u6b21\u5b9e\u9a8c\u4e2d\u662f\u5b8c\u5168\u76f8\u53cd\u7684\uff0c\u5373\u4e0b\u56fe\u4e2d\u6846\u6846\u6846\u8d77\u6765\u7684\u90e8\u5206\u3002<\/p>\n\n\n\n<figure class=\"wp-block-image\"><img loading=\"lazy\" decoding=\"async\" decoding=\"async\" src=\"https:\/\/eamonimgbed.oss-cn-beijing.aliyuncs.com\/img\/202304052023340.webp\" alt=\"WB \u62f7\u8d1d 3\" \/><\/figure>\n\n\n\n<h2 class=\"wp-block-heading\">\u4fe1\u53f7\u6574\u4f53\u504f\u5f31\u65f6\u7684\u7ed3\u679c\u4e0d\u53ef\u4fe1<\/h2>\n\n\n\n<p>\u4e4b\u524d\u6211\u7528\u8fc7\u7684\u5185\u53c2\u6709beta-actin\u548cbeta-tubulin\uff0c\u8fd9\u6b21\u6253\u5361\u4e86HSP90\u3002<\/p>\n\n\n\n<p>\u6211\u8fd9\u4e2a\u6708\u7b2c\u4e00\u6b21\u8dd1\u7684\u65f6\u5019\uff0c\u9047\u5230\u4e86actin\u4e0d\u9f50\u7684\u95ee\u9898\uff0c\u86cb\u767d\u6837\u54c1\u5df2\u7ecf\u7528BCA\u6cd5\u5b9a\u91cf\u8fc7\u4e86\u3002\u4f46\u662f\u9047\u5230\u4e86\u540c\u4e00\u62e8\u6837\u54c1\u3001\u76f8\u540c\u7684\u4e0a\u6837\u91cf\u7684\u4e09\u6b21\u5e73\u884c\u91cd\u590d\u4e2dactin\u6761\u5e26\u7684\u76f8\u5bf9\u4eae\u5ea6\u5747\u4e0d\u4e00\u81f4\u7684\u95ee\u9898\u3002\u540e\u6765\u6362\u4e86\u4e00\u4e2a\u522b\u7684\u724c\u5b50\u7684\u6297\u4f53\uff0c\u5c31\u6bcf\u6b21\u90fd\u5929\u7136\u5185\u53c2\u4e00\u81f4\u4e86\u3002\u53ef\u80fd\u662f\u7b2c\u4e00\u6b21\u7528\u7684\u90a3\u4e2a\u6297\u4f53\u8981\u4e48\u56de\u6536\u6b21\u6570\u8fc7\u591a\u3001\u8981\u4e48\u672c\u8eab\u5c31\u4e0d\u884c\u3002\u6d6a\u8d39\u4e86\u6211\u6574\u6574\u4e09\u6b21\u5b8c\u6574\u7684WB\u8017\u8d39\u7684\u65f6\u95f4\u3002<\/p>\n\n\n\n<p>\u4e4b\u540e\u7684WB\u6211\u90fd\u76f4\u63a5\u7528\u4e24\u4e2a\u5185\u53c2\u53bb\u8dd1\u4e86\uff0c\u540c\u65f6\u5b75anti-actin\u548canti-HSP90\u3002\u56de\u5934\u60f3\u60f3\uff0c\u5bf9\u4e8eactin\u8fd9\u79cd\u5546\u4e1a\u5316\u6297\u4f53\uff0c\u4e14\u86cb\u767d\u672c\u8eab\u8868\u8fbe\u91cf\u9ad8\uff0c\u5e94\u8be51s\u5c31\u80fd\u66dd\u51fa\u5f88\u4eae\u7684\u6761\u5e26\u624d\u5bf9\uff0c\u5f53\u6211\u7b2c\u4e00\u6b21WB\u663e\u5f71\u65f6\u53d1\u73b0\u66dd\u514930s\u4ee5\u4e0a\u624d\u51fa\u73b0\u90a3\u6837\u5fae\u5f31\u7684\u6761\u5e26\u65f6\uff0c\u5c31\u5e94\u8be5\u7acb\u523b\u610f\u8bc6\u5230\u6297\u4f53\u51fa\u95ee\u9898\u4e86\uff0c\u800c\u4e0d\u662f\u7528\u8fd9\u4e2a\u6839\u672c\u4e0d\u53ef\u4fe1\u7684\u6570\u636e\u53bb\u65e0\u610f\u4e49\u5730\u8c03\u6574\u4e0a\u6837\u91cf\u3002<\/p>\n\n\n\n<figure class=\"wp-block-image\"><img loading=\"lazy\" decoding=\"async\" decoding=\"async\" src=\"https:\/\/eamonimgbed.oss-cn-beijing.aliyuncs.com\/img\/202304052049880.png\" alt=\"actin\" \/><\/figure>\n\n\n\n<p>\u53e6\u5916\uff0c\u6211\u503e\u5411\u4e8e\u8ba4\u4e3a\uff0c\u5f53WB\u4fe1\u53f7\u6574\u4f53\u975e\u5e38\u4f4e\u65f6\uff0c\u6bd4\u8f83\u4e24\u4e2a\u6cf3\u9053\u7684\u76f8\u5bf9\u6761\u5e26\u4eae\u5ea6\u5dee\u5f02\u662f\u6781\u5176\u4e0d\u51c6\u786e\u7684\uff0c\u5982\u7b2c\u4e00\u5f20\u56fe\u4e2dProtein A\u88ab\u6846\u8d77\u6765\u7684\u6761\u5e26\uff0c\u5728\u4e24\u6b21\u91cd\u590d\u5b9e\u9a8c\u4e2d\u7684\u76f8\u5bf9\u4eae\u5ea6\u5b8c\u5168\u76f8\u53cd\u3002<\/p>\n\n\n\n<h2 class=\"wp-block-heading\">\u6297\u4f53\u9009\u62e9\u95ee\u9898<\/h2>\n\n\n\n<p>\u6297\u4f53\u4e0d\u597d\u4e0d\u8981\u627e\u81ea\u5df1\u7684\u95ee\u9898\uff0c\u4e0a\u9762\u90a3\u4e2aactin\u662f\u4e00\u4e2a\u4f8b\u5b50\uff0c\u5bf9\u4e8e\u4e0d\u5e38\u7528\u86cb\u767d\uff0c\u6297\u4f53\u672c\u8eab\u5c31\u4e0d\u884c\u7684\u53ef\u80fd\u5f88\u5927\u3002\u6bd4\u5982Protein I\u6709IS\u548cIU\u4e24\u79cd\u5f62\u5f0f\uff0c\u7528\u6297\u4f53\u4e00\u5c31\u53ea\u80fd\u5f97\u5230IU\u5f62\u5f0f\uff0c\u5f97\u4e0d\u5230IS\u5f62\u5f0f\uff0c\u800c\u7528\u6297\u4f53\u4e8c\u53ef\u4ee5\u5f97\u5230IS\u5f62\u5f0f\uff0c\u53ef\u662fIU\u6761\u5e26\u5c31\u8981\u5f31\u5f97\u591a\u3002\u5173\u952e\u662f\uff0c\u8fd9\u4e24\u5bb6\u6297\u4f53\u7684\u8bf4\u660e\u4e66\u90fd\u5199\u4e86\u80fd\u7838\u5230\u4e24\u79cd\u5f62\u5f0f\u7684\u6761\u5e26\u3002<\/p>\n\n\n\n<figure class=\"wp-block-image\"><img loading=\"lazy\" decoding=\"async\" decoding=\"async\" src=\"https:\/\/eamonimgbed.oss-cn-beijing.aliyuncs.com\/img\/202304052140034.png\" alt=\"image-20230405214036153\" \/><\/figure>\n\n\n\n<h2 class=\"wp-block-heading\">\u663e\u8272\u6db2\u95ee\u9898<\/h2>\n\n\n\n<p>\u6211\u6709\u6b21\u66dd\u51fa\u6765\u8fd9\u6837\u4e00\u5f20\u56fe\uff0c\u6700\u521d\u662f\u4ee5\u4e3a\u6ca1\u6709\u6d17\u5e72\u51c0\u3002\u6bd4\u5982\u6d17\u819c\u65f6TBST\u52a0\u7684\u592a\u5c11\uff0c\u6240\u4ee5\u819c\u4e2d\u95f4\u90e8\u5206\u6d17\u5f97\u4e0d\u5145\u5206\uff0c\u5bfc\u81f4\u4e2d\u95f4\u6bd4\u8f83\u810f\u800c\u4e24\u4fa7\u5e72\u51c0\u3002\u540e\u6765\u6211\u53d1\u73b0\u5176\u5b9e\u53ea\u662f\u6211\u7684\u663e\u8272\u6db2\u8986\u76d6\u7684\u4e0d\u591f\u5747\u5300\u3002\u8fd9\u4e2a\u5c0f\u7ec6\u8282\u592a\u91cd\u8981\u4e86\uff0c\u5b83\u53ef\u80fd\u76f4\u63a5\u5f71\u54cd\u4e0d\u540c\u6cf3\u9053\u4e4b\u95f4\u7684\u6761\u5e26\u4eae\u5ea6\u5dee\u5f02\u3002\u4ee5\u540e\u52a0\u663e\u8272\u6db2\u65f6\uff0c\u4e00\u5b9a\u8981\u5de6\u53f3\u6643\u52a8\u6216\u7528\u5e73\u677f\u538b\u7247\u7b49\u65b9\u6cd5\u628a\u663e\u8272\u6db2\u52a0\u5747\u5300\u3002<\/p>\n\n\n\n<figure class=\"wp-block-image\"><img loading=\"lazy\" decoding=\"async\" decoding=\"async\" src=\"https:\/\/eamonimgbed.oss-cn-beijing.aliyuncs.com\/img\/202304052136347.png\" alt=\"\u663e\u5f71\u6db2\u52a0\u5c11 \u62f7\u8d1d\" \/><\/figure>\n\n\n\n<h2 class=\"wp-block-heading\">\u66dd\u5149\u65f6\u95f4\u957f\u77ed\u95ee\u9898<\/h2>\n\n\n\n<p>\u4f60\u770b\u8fd9\u4e2aHSP90\uff0c\u5149\u770b\u66dd\u51491s\uff0c3s\uff0c10s\u7684\u56fe\uff0c\u4e00\u770b\u5c31\u5185\u53c2\u4e0d\u9f50\uff0c\u8981\u91cd\u65b0\u505a\u3002\u4f46\u662f\u518d\u4e00\u770b\u66dd\u514930s\u7684\uff0c\u597d\u50cf\u4e5f\u4e0d\u662f\u90a3\u4e48\u4e0d\u9f50\u2026\u2026\u4f60\u8bf4\u8fd9\u7b97\u4e0d\u7b97\u9020\u5047\u5462\uff1f<\/p>\n\n\n\n<figure class=\"wp-block-image\"><img loading=\"lazy\" decoding=\"async\" decoding=\"async\" src=\"https:\/\/eamonimgbed.oss-cn-beijing.aliyuncs.com\/img\/202304052151644.webp\" alt=\"\u66dd\u5149\u65f6\u95f4 \u62f7\u8d1d\" \/><\/figure>\n\n\n\n<h2 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