{"id":2984,"date":"2024-06-19T21:01:29","date_gmt":"2024-06-19T13:01:29","guid":{"rendered":"https:\/\/opentrons.com.cn\/?post_type=knowledge2&#038;p=2984"},"modified":"2024-09-02T18:57:30","modified_gmt":"2024-09-02T10:57:30","slug":"thermo-fisher-dynabeads-jxcdmy","status":"publish","type":"post","link":"https:\/\/opentrons.com.cn\/news\/thermo-fisher-dynabeads-jxcdmy\/","title":{"rendered":"\u5728OT-2\u4e0a\u4f7f\u7528Thermo Fisher Dynabeads \u8fdb\u884c\u514d\u75ab\u6c89\u6dc0"},"content":{"rendered":"\n<p><strong>\u4f5c\u8005<\/strong><\/p>\n\n\n\n<p>Boren \u00a0Lin,PhD1,Kinnari \u00a0Watson,PhDl 1Opentrons\u00a0\u00a0Labworks,Inc.<\/p>\n\n\n\n<p><strong>\u6458\u8981<\/strong><br>\u672c\u6b21\u7814\u7a76\u5728Opentrons OT-2\u81ea\u52a8\u5316\u79fb\u6db2\u5de5\u4f5c\u7ad9\u4e0a\u8fdb\u884c\u4e86\u81ea \u52a8\u5316\u7684\u78c1\u73e0\u6cd5\u514d\u75ab\u6c89\u6dc0\u6d4b\u8bd5\uff0c\u5b9e\u9a8c\u6d41\u7a0b\u53ef\u5904\u7406\u591a\u8fbe96\u4e2a\u6837 \u54c1\uff0c\u5e76\u6700\u5927\u9650\u5ea6\u5730\u51cf\u5c11\u4eba\u5de5\u64cd\u4f5c\u65f6\u95f4\u3002\u6211\u4eec\u7528\u88c2\u89e3\u7f13\u51b2\u6db2 \u7a00\u91ca\u91cd\u7ec4 GAPDH \u86cb\u767d\u6216\u8005\u76f4\u63a5\u4f7f\u7528 HEp-2 \u7ec6\u80de\u88c2\u89e3\u4ea7\u7269 \u8fdb\u884c\u6837\u672c\u5236\u5907\uff0c\u5e76\u5076\u8054\u86cb\u767dG \u6216\u8005\u86cb\u767dA, \u7136\u540e\u5728 Thermo Fisher Dynabeads \u4e0a\u8fdb\u884c\u514d\u75ab\u6c89\u6dc0\u5b9e\u9a8c(\u86cb\u767dG\u548c\u86cb\u767dA \u53ef\u4e0e\u53ef\u6eb6\u6027\u6297\u4f53\u7ed3\u5408\uff0c\u8fdb\u800c\u4fc3\u8fdb\u968f\u540e\u4e0e\u76ee\u6807\u86cb\u767d GAPDH \u7684 \u7ed3\u5408)\u3002\u86cb\u767d\u8d28\u5206\u6790\u7ed3\u679c\u663e\u793a\uff0c\u76ee\u6807\u86cb\u767d\u8d28\u6210\u529f\u5730\u4ece\u6837\u54c1\u6eb6 \u6db2\u4e2d\u63d0\u53d6\u51fa\u6765\uff0c\u5e76\u7b26\u5408\u9884\u671f\u6548\u679c\u7684\u4e00\u81f4\u6027\uff0c\u8bc1\u660e\u4e86 OT-2 \u5177\u5907\u81ea\u52a8\u5316\u8fdb\u884c\u514d\u75ab\u6c89\u6dc0\u5b9e\u9a8c\u7684\u80fd\u529b\u3002<\/p>\n\n\n\n<p><strong>\u7b80\u4ecb<\/strong><br>\u514d\u75ab\u6c89\u6dc0\u662f\u4e00\u79cd\u5e38\u89c1\u7684\u5b9e\u9a8c\u65b9\u6cd5\uff0c\u5229\u7528\u6297\u4f53\u5728\u5206\u5b50\u6df7\u5408\u7269 \u4e2d\u6355\u83b7\u76ee\u6807\u86cb\u767d\u8d28\u3002\u6297\u4f53\u53ef\u4ee5\u9884\u5148\u8fde\u63a5\u5230\u56fa\u76f8\u57fa\u8d28(\u5982\u743c \u8102\u7cd6\u6811\u8102\u6216\u78c1\u73e0)\u4e0a\uff0c\u7136\u540e\u4e0e\u76ee\u6807\u86cb\u767d\u8d28\u63a5\u89e6\uff0c\u6216\u8005\u76f4\u63a5 \u4e0e\u6837\u54c1\u4e00\u8d77\u52a0\u5165\u76f8\u5e94\u7684\u56fa\u76f8\u57fa\u8d28\u3002\u7531\u4e8e\u5b75\u80b2\u65f6\u95f4\u53d6\u51b3\u4e8e\u6297 \u4f53\u4e0e\u76ee\u6807\u86cb\u767d\u8d28\u7684\u4eb2\u548c\u529b\uff0c\u4e0d\u540c\u7684\u76ee\u6807\u6355\u83b7\u86cb\u767d\u6240\u9700\u7684\u5b75 \u80b2\u65f6\u95f4\u4e5f\u4f1a\u4e0d\u4e00\u6837\u3002\u4f7f\u7528\u78c1\u73e0\u63d0\u53d6\uff0c\u6297\u4f53-\u86cb\u767d\u8d28-\u78c1\u73e0\u590d\u5408\u7269\u4f1a\u901a\u8fc7\u79bb\u5fc3\u6216\u78c1\u529b\u7684\u65b9\u5f0f\u8fdb\u884c\u6c89\u6dc0\u6536\u96c6\u3002<\/p>\n\n\n\n<p>DynabeadsM(Thermo Fisher Scientific,USA) \u662f\u4e00\u6b3e\u53ef \u4ee5\u6839\u636e\u9700\u6c42\u8c03\u6574\u4ee5\u83b7\u5f97\u4e0d\u540c\u529f\u80fd\u7684\u8d85\u987a\u78c1\u73e0\u3002\u4f8b\u5982\uff0c\u91cd\u7ec4\u7684 \u86cb\u767dG\u6216\u86cb\u767dA\u53ef\u4ee5\u5171\u4ef7\u5730\u7ed3\u5408\u5230\u78c1\u73e0\u8868\u9762\u3002\u86cb\u767dG\u548c\u86cb\u767dA \u90fd\u662f\u7ec6\u83cc\u86cb\u767d\uff0c\u5bf9\u5355\u514b\u9686\u6216\u591a\u514b\u9686 IgG \u578b\u6297\u4f53\u7684 Fc \u533a\u57df\u8868\u73b0 \u51fa\u9ad8\u4eb2\u548c\u529b\u3002DynabeadsTM- \u86cb\u767dG \u6216 DynabeadsTM- \u86cb\u767d A\u53ef\u4f5c\u4e3a\u6297\u4f53\u7684\u56fa\u76f8\u652f\u6301\u6750\u6599\u3002Opentrons \u4ee5 DynabeadsM\u4e3a\u57fa\u7840\u8bbe\u8ba1\u4e86\u9ad8\u6548\u81ea\u52a8\u5316\u7684\u5b9e\u9a8c\u7a0b\u5e8f\u8fdb\u884c\u514d\u75ab\u6c89\u6dc0\u5b9e\u9a8c\uff0c\u6bcf\u6b21\u5b9e\u9a8c\u53ef\u5904\u7406\u6700\u591a96\u4e2a\u6837\u54c1\uff0c\u6837\u54c1\u4e4b\u95f4\u5177\u6709\u826f\u597d\u7684\u91cd\u590d\u6027\u3002<\/p>\n\n\n\n<p><strong>\u6750\u6599\u548c\u65b9\u6cd5<\/strong><br>\u5b9e\u9a8c\u6b65\u9aa4\u5305\u62ec\u4ee5\u4e0b\u5185\u5bb9\uff1a<br>\u7b2c\u4e00\u90e8\u5206\u662f\u78c1\u73e0\u3001\u6297\u4f53\u548c\u6837\u54c1\u7684\u6df7\u5408\u5236\u5907\uff1b\u7531\u7528\u6237\u786e\u5b9a\u7684\u6297\u4f53\/\u76ee\u6807\u86cb\u767d\u7684\u5b75\u80b2\u65f6\u95f4\uff1b<br>\u7b2c\u4e8c\u90e8\u5206\u662f\u6d17\u6da4\u548c\u6d17\u8131\u6b65\u9aa4(\u56fe1)\u3002<br>\u7b2c\u4e00\u90e8\u5206\u548c\u7b2c\u4e8c\u90e8\u5206\u5728 OT-2 \u4e0a\u8fdb\u884c\uff0c\u4f7f\u7528\u78c1\u73e0\u7eaf\u5316\u6a21\u5757\u5c06\u78c1 \u73e0\u4ece\u6eb6\u6db2\u4e2d\u5206\u79bb\u51fa\u6765\uff0c\u5e76\u4f7f\u7528\u6e29\u63a7\u6a21\u5757\u5236\u5907\u53d8\u6027\u86cb\u767d\u7684\u6d17\u8131\u6db2\u8fdb\u884c SDS-PAGE\u3002 \u5b75\u80b2\u8fc7\u7a0b\u5728\u70ed\u632f\u8361\u4eea(\u9009\u914d)\u4e0a\u8fdb\u884c\u3002<br>\u514d\u75ab\u6c89\u6dc0\u5b9e\u9a8c\u7684\u7b2c\u4e00\u90e8\u5206\u548c\u7b2c\u4e8c\u90e8\u5206\u7684 OT-2 \u88c5\u8f7d\u5e03\u5c40\u5982\u56fe2 \u6240\u793a\u3002\u8fd9\u4e2a\u6d41\u7a0b\u65e8\u5728\u5c06\u8bd5\u5242\u76f4\u63a5\u4ece15 mL \u79bb\u5fc3\u7ba1\u8f6c\u79fb\u5230\u5de5\u4f5c\u677f\u4e0a\u8fdb\u884c\u78c1\u73e0\u5206\u79bb\u3002<br>\u53e6\u5916\uff0c\u5728 OT-2 \u4e0a\u4f7f\u7528\u81ea\u52a8\u5316\u7a0b\u5e8f\u9884\u586b\u5145\u8bd5\u5242\u677f\uff0c\u53ef\u4ee5\u51cf\u5c11\u5b9e \u9a8c\u65f6\u95f4\u3002\u6297\u4f53\u4e0e\u76ee\u6807\u86cb\u767d\u7684\u7ed3\u5408\u662f\u514d\u75ab\u6c89\u6dc0\u6210\u529f\u7684\u6700\u5173\u952e\u6b65 \u9aa4\uff0c\u8fd9\u53d6\u51b3\u4e8e\u6297\u4f53\u7684\u4eb2\u548c\u529b\u3002\u5bf9\u4e8e\u7ed3\u5408\u4eb2\u548c\u529b\u4f4e\u6216\u4e30\u5ea6\u8f83\u4f4e\u7684\u6297\u4f53\uff0c\u5efa\u8bae\u5728\u6837\u54c1\u6eb6\u6db2\u4e2d\u8fdb\u884c\u9884\u5b75\u80b2\u3002<\/p>\n\n\n\n<figure class=\"wp-block-image size-full\"><img decoding=\"async\" width=\"650\" height=\"273\" src=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-59.png\" alt=\"\u86cb\u767d\u7eaf\u5316\u7684\u4e09\u4e2a\u4e3b\u8981\u6b65\u9aa4\u7684\u6982\u8ff0\" class=\"wp-image-2985\" srcset=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-59.png 650w, https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-59-300x126.png 300w, https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-59-576x242.png 576w\" sizes=\"(max-width: 650px) 100vw, 650px\" \/><\/figure>\n\n\n\n<figure class=\"wp-block-image size-full is-resized\"><img decoding=\"async\" width=\"707\" height=\"425\" src=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-60.png\" alt=\"\u78c1\u73e0\u7eaf\u5316\u6a21\u5757\" class=\"wp-image-2986\" style=\"width:481px;height:auto\" srcset=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-60.png 707w, https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-60-300x180.png 300w, https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-60-576x346.png 576w\" sizes=\"(max-width: 707px) 100vw, 707px\" \/><\/figure>\n\n\n\n<figure class=\"wp-block-image size-full is-resized\"><img decoding=\"async\" width=\"690\" height=\"429\" src=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-61.png\" alt=\"\u78c1\u73e0\u7eaf\u5316\u6a21\u5757\" class=\"wp-image-2987\" style=\"width:453px;height:auto\" srcset=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-61.png 690w, https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-61-300x187.png 300w, https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-61-576x358.png 576w\" sizes=\"(max-width: 690px) 100vw, 690px\" \/><\/figure>\n\n\n\n<figure class=\"wp-block-image size-full is-resized\"><img decoding=\"async\" width=\"732\" height=\"453\" src=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-62.png\" alt=\"\u8017\u6750\u548c\u6a21\u5757\u7532\u677f\u6446\u653e\u793a\u610f\u56fe\" class=\"wp-image-2988\" style=\"width:444px;height:auto\" srcset=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-62.png 732w, https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-62-300x186.png 300w, https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-62-576x356.png 576w\" sizes=\"(max-width: 732px) 100vw, 732px\" \/><\/figure>\n\n\n\n<figure class=\"wp-block-image size-full\"><img decoding=\"async\" width=\"829\" height=\"380\" src=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-63.png\" alt=\"\u8bd5\u7ba1\u8bd5\u5242\u793a\u610f\u56fe\" class=\"wp-image-2989\" srcset=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-63.png 829w, https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-63-300x138.png 300w, https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-63-768x352.png 768w, https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-63-576x264.png 576w\" sizes=\"(max-width: 829px) 100vw, 829px\" \/><figcaption class=\"wp-element-caption\">\u56fe4:\u8bd5\u7ba1\u91cc\u7684\u8bd5\u5242\u548c\u5b54\u677f\u91cc\u7684\u8bd5\u5242\u6240\u9700\u7684\u8fd0\u884c\u65f6\u95f4\u548c\u76d2\u88c5\u5438\u5934\u6570\u91cf\u3002<\/figcaption><\/figure>\n\n\n\n<p><strong>\u7ed3\u679c<\/strong><br>\u5154\u5b50IgG \u5bf9 \u86cb \u767dG \u548c\u86cb\u767dA \u90fd\u5177\u6709\u9ad8\u4eb2\u548c\u6027\uff0c\u4e3a\u4e86\u786e\u8ba4\u6297\u4f53\u4e0e DynabeadsM \u7684\u76f8\u4e92\u4f5c\u7528\uff0c\u6211\u4eec\u4f7f\u7528\u4e86\u6297 GAPDH \u7684\u514d\u591a\u514b\u9686 gG \u6297\u4f53(ProteintechRosemont,IL,USA) \u3002 \u5728 OT-2 \u4e0a\uff0c\u5c06 50\u03bcL \u7684\u78c1\u73e0\u60ac\u6db2\u4e0e\u542b\u67092.5\u03bcg GAPDH \u6297\u4f53\u7684200\u03bcL PBS \u6df7 \u5408\uff0c\u7136\u540e\u5ba4\u6e29\u4e0b\u4e8e\u632f\u8361\u5668\u4e0a800 rpm \u6df7\u530010\u5206\u949f\u3002\u968f\u540e\uff0c\u5728 OT-2\u4e0a\u75280 . 1% Tween 20 \u7684\u78f7\u9178\u76d0\u7f13\u51b2\u751f\u7406\u76d0\u6c34 (PBS-T) \u6d17\u6da43\u6b21\u300270\u00b0\u2103\u52a0\u70ed10\u5206\u949f\u3001\u4f7f\u7528Laemmli \u6837\u54c1\u7f13\u51b2\u6db2 (Bio-Rad,USA) \u6d17\u8131\u7ed3\u5408\u7684 GAPDH \u6297\u4f53\u3002\u7136\u540e\uff0c\u5bf9\u5176\u8fdb \u884cSDS-PAGE \u5206\u79bb\uff0c\u518d\u7528\u8fd1\u7ea2\u5916\u53d1\u5149\u67d3\u6599 (IRDye 680RD)\u5076 \u8054\u7684\u5c71\u7f8a\u6297\u5154 IgG \u6297\u4f53 (LI-COR Biosciences,USA)\u8fdb\u884c\u63a2\u9488\u63a2\u6d4b\u3002Western Blot \u5206\u6790\u7ed3\u679c\u663e\u793a\uff0c\u5f53\u8fdb\u884c\u56db\u6b21\u91cd\u590d\u64cd\u4f5c\u65f6\uff0c\u65e0\u8bba\u662f\u86cb\u767dG \u8fd8\u662f\u86cb\u767dA \u78c1\u73e0\u90fd\u80fd\u591f\u4e0e GAPDH \u6297\u4f53\u7ed3\u5408(\u56fe5A), \u4e14\u7ed3\u5408\u5177\u5907\u53ef\u91cd\u590d\u6027\u3002<br>\u4e3a\u4e86\u8fdb\u884c GAPDH \u7684\u514d\u75ab\u6c89\u6dc0\uff0c\u6211\u4eec\u5728 OT-2 \u4e0a\u4f7f\u7528\u542b\u6709\u91cd\u7ec4 \u4eba GAPDH \u86cb \u767d(rhGAPDH,Thermo Fisher Scientific, USA) \u7684\u6837\u54c1\u8fdb\u884c\u514d\u75ab\u6c89\u6dc0\u5904\u7406\u3002\u540c\u65f6\u4f7f\u7528\u7ed3\u5408\u4e86 GAPDH \u6297 \u4f53\u7684 DynabeadsTM-\u86cb\u767dG\u548c\u86cb\u767dA\u7684 GAPDH\u6297\u4f53\u8fdb\u884c\u6d4b\u8bd5\u3002 Western Blot\u7ed3\u679c\u663e\u793a\u51faGAPDH \u7684\u5b58\u5728\uff0c\u9a8c\u8bc1\u4e86OT-2 \u5177\u5907\u8fd0\u884c\u81ea\u52a8\u5316\u514d\u75ab\u6c89\u6dc0\u7684\u80fd\u529b(\u56fe5B)\u3002<br>\u8be5\u65b9\u6cd5\u8fd8\u9488\u5bf9\u57f9\u517b\u7ec6\u80de\u4e2d\u5185\u6e90\u6027 GAPDH \u8fdb\u884c\u4e86\u81ea\u52a8\u5316\u514d\u75ab\u6c89\u6dc0\u8bc4\u4f30\u3002HEp-2 \u7ec6\u80de\u5728\u542b\u670910%\u80ce\u725b\u8840\u6e05\u548c1%\u9752\u9709\u7d20-\u94fe\u9709\u7d20\u7684 GibcoTM\u6700\u4f4e\u57fa\u7840\u57f9\u517b\u57fa(Thermo Fisher Scientific,USA) \u4e2d\u57f9\u517b\uff0c\u57285% CO2 \u548c37\u00b0\u2103\u6761\u4ef6\u4e0b\u8fdb\u884c\u3002\u4e3a\u4e86\u51c6\u5907\u6d4b\u8bd5\u7684\u7ec6\u80de\u88c2\u89e3\u6db2\uff0c\u6536\u96c6\u4e864 x106 \u4e2a HEp-2 \u7ec6\u80de\uff0c\u5e76\u5728200\u03bcLPierceTM IP \u7684\u88c2\u89e3\u7f13\u51b2\u6db2\u4e2d\u8fdb\u884c\u88c2\u89e3\uff0c\u8be5\u7f13\u51b2\u6db2\u4e2d\u8fd8\u52a0\u5165\u4e86Thermo Fisher Scientific \u63d0\u4f9b\u7684\u86cb\u767d\u9176\u6291\u5236\u5242\u6df7\u5408\u7269\u3002\u4f7f\u7528DynabeadsTM- \u86cb\u767dA\u5728 OT-2 \u5e73\u53f0\u4e0a\u8fdb\u884c\u514d\u75ab\u6c89\u6dc0\u5b9e\u9a8c\uff0cOT-2\u79fb\u6db2\u5e73\u53f0\u642d\u8f7d\u4e86\u78c1\u73e0\u7eaf\u5316\u6a21\u5757\u548c\u6e29\u63a7\u6a21\u5757\u3002\u4e3a\u4e86\u8ba9\u6297\u4f53\u4e0e\u76ee\u6807\u86cb\u767d\u7ed3\u5408\u6709\u8db3\u591f\u7684\u65f6\u95f4\uff0c\u5c06\u6df7\u5408\u7269\u653e\u7f6e\u4e8e4\u00b0\u2103\u70ed\u632f\u8361\u4eea\u4e0a\uff0c\u5728800 rpm\u7684\u8f6c\u901f\u4e0b\u5b75\u80b2\u8fc7\u591c\u3002\u6700\u540e\u901a\u8fc7Western Blot 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