{"id":7975,"date":"2025-09-12T13:36:39","date_gmt":"2025-09-12T13:36:39","guid":{"rendered":"https:\/\/nanomicronspheres.com\/protocolo-de-beads-magneticas-para-captura-de-gfp1-materiais-necessarios-beads-magneticas-especificas-para-gfp-solucao-de-lise-celular-tampao-de-lavagem-tampao-de-eluicao\/"},"modified":"2025-09-12T13:36:39","modified_gmt":"2025-09-12T13:36:39","slug":"protocolo-de-beads-magneticas-para-captura-de-gfp1-materiais-necessarios-beads-magneticas-especificas-para-gfp-solucao-de-lise-celular-tampao-de-lavagem-tampao-de-eluicao","status":"publish","type":"post","link":"https:\/\/nanomicronspheres.com\/zh\/protocolo-de-beads-magneticas-para-captura-de-gfp1-materiais-necessarios-beads-magneticas-especificas-para-gfp-solucao-de-lise-celular-tampao-de-lavagem-tampao-de-eluicao\/","title":{"rendered":"Otimizando Seus \u7ed3\u679c\uff1aUm Guia Passo a Passo para o Protocolo de Beads Magn\u00e9ticos GFP Trap"},"content":{"rendered":"<p>\u6ca1\u6709\u751f\u7269\u5b66\u5206\u5b50\uff0c\u78c1\u73e0\u534f\u8bae GFP \u9677\u9631\u662f\u5206\u79bb\u548c\u7eaf\u5316\u86cb\u767d\u8d28\u7684\u57fa\u672c\u6280\u672f\uff0c\u5305\u62ec\u7eff\u8272\u8367\u5149\u86cb\u767d (GFP)\u3002\u6539\u8fdb\u7684\u65b9\u6cd5\u662f\u7eaf\u5316\u86cb\u767d\u8d28\u4f5c\u4e3a\u8fde\u63a5\u78c1\u73e0\u7684\u5c5e\u6027\uff0c\u4ee5\u5bf9\u6297 GFP \u7684\u7279\u6b8a\u60c5\u51b5\u3002\u8be5\u534f\u8bae\u7b80\u5316\u4e86\u751f\u7269\u590d\u5408\u4f53\u7684\u5904\u7406\u8fc7\u7a0b\uff0c\u5141\u8bb8\u7814\u7a76\u4eba\u5458\u4ee5\u6709\u6548\u7684\u65b9\u5f0f\u7814\u7a76\u86cb\u767d\u8d28\u7684\u5174\u8da3\uff0c\u5e76\u4e0e\u7ec6\u80de\u4e2d\u7684\u6210\u5206\u4e00\u8d77\u4f7f\u7528\u3002<\/p>\n<p>\u78c1\u73e0\u534f\u8bae GFP Trap \u53ef\u4ee5\u6709\u6548\u7eaf\u5316\uff0c\u5e76\u4fdd\u8bc1\u7279\u5b9a\u7684\u60c5\u51b5\u548c\u51cf\u5c11\u7279\u5b9a\u7684\u8fde\u63a5\u3002\u8fd9\u662f\u5173\u4e8e\u9ad8\u7ea7\u86cb\u767d\u8d28\u7684\u989d\u5916\u4fe1\u606f\uff0c\u662f\u540e\u90e8\u5206\u6790\u7684\u672c\u8d28\uff0c\u4e5f\u662f\u529f\u80fd\u548c\u7814\u7a76\u7684\u57fa\u7840\u3002\u8fd9\u662f\u79d1\u5b66\u6280\u672f\u7814\u7a76\u7684\u5fc5\u8981\u6761\u4ef6\u548c\u86cb\u767d\u8d28\u7eaf\u5316\u7684\u5fc5\u8981\u6761\u4ef6\uff0c\u78c1\u73e0 GFP \u9677\u9631\u534f\u8bae\u662f\u86cb\u767d\u8d28\u7814\u7a76\u7684\u91cd\u8981\u6b66\u5668\u5e93\u3002<\/p>\n<p>\u5728\u6b64\u8fc7\u7a0b\u4e2d\uff0c\u6211\u4eec\u5c06\u4e86\u89e3\u5b9e\u9a8c\u7ed3\u679c\uff0c\u5e76\u6df1\u5165\u4e86\u89e3\u5404\u79cd\u751f\u7269\u5b66\u8fc7\u7a0b\u4e2d\u7684\u86cb\u767d\u8d28\u76f8\u4e92\u4f5c\u7528\u3001\u529f\u80fd\u548c\u86cb\u767d\u8d28\u3002<\/p>\n<h2>Como o Protocolo de Esferas Magn\u00e9ticas GFP Trap Melhora \u548c Purifica\u00e7\u00e3o de Prote\u00ednas<\/h2>\n<p>\u86cb\u767d\u8d28\u7684\u7eaf\u5316\u901a\u8fc7\u751f\u7269\u5206\u6790\u65b9\u6cd5\u7684\u6279\u5224\uff0c\u5141\u8bb8\u5bf9\u7279\u5b9a\u86cb\u767d\u8d28\u8fdb\u884c\u8be6\u7ec6\u7814\u7a76\u3002\u4f5c\u4e3a\u591a\u79cd\u6280\u672f\u7684\u4e00\u90e8\u5206\uff0cEsferas Magn\u00e9ticas GFP Trap \u534f\u8bae\u662f\u4e00\u79cd\u6709\u6548\u7684\u86cb\u767d\u8d28\u7eaf\u5316\u65b9\u6cd5\u548c\u5b9e\u8df5\u65b9\u6cd5\u3002\u8be5\u534f\u8bae\u5df2\u88ab\u8bc1\u5b9e\u4e3a Prote\u00edna Fluorescente Verde (GFP) \u7684\u5c5e\u6027\uff0c\u7528\u4e8e\u5206\u79bb\u86cb\u767d\u8d28\uff0c\u5e76\u6709\u52a9\u4e8e\u7eaf\u5316\u8fc7\u7a0b\u7684\u8d28\u91cf\u3002<\/p>\n<h3>Entendendo \u9970\u6f14 Esferas Magn\u00e9ticas GFP Trap<\/h3>\n<p>\u4f5c\u4e3a Esferas Magn\u00e9ticas GFP \u9677\u9631\uff0c\u5b83\u53ef\u4ee5\u6355\u83b7 GFP \u7684\u7279\u5b9a\u86cb\u767d\u8d28\u3002 Essas esferas fornecem uma \u5e73\u53f0robusta para isolar prote\u00ednas de milsuras Complexas\uff0ccomo lisados cellulares\u3002\u7531\u4e8e\u4e0e GFP \u7684\u53cd\u4f53\u91cd\u7ec4\u548c\u8fde\u63a5\u6709\u5173\uff0c\u56e0\u6b64\u5141\u8bb8\u5c06 GFP \u4e0e\u975e\u6d3b\u6027\u86cb\u767d\u8d28\u548c\u6c61\u67d3\u7269\u5206\u5f00\u3002\u6211\u4eec\u63a2\u7d22\u4e86\u4e00\u7cfb\u5217\u7684\u4ea4\u4e92\u65b9\u5f0f\uff0c\u4f7f\u7814\u7a76\u4eba\u5458\u80fd\u591f\u66f4\u7b80\u5355\u5730\u4e86\u89e3\u51c0\u5316\u534f\u8bae\u7684\u610f\u4e49\u3002<\/p>\n<h3>\u4f7f\u7528 GFP \u9677\u9631\u534f\u8bae\u7684\u597d\u5904<\/h3>\n<p>Empregar o \u534f\u8bae de Esferas Magn\u00e9ticas GFP Trap oferece in\u00famerasvantagens, \u9f99\u5377\u98ce-o uma escolha prencial para muitos pesquisadores:<\/p>\n<ul>\n<li><strong>\u9009\u62e9:<\/strong> O \u534f\u8bae\u4fdd\u8bc1\u4e86\u7279\u5b9a\u7684\u60c5\u51b5\uff0c\u6355\u83b7\u4e86 GFP \u7684\u86cb\u767d\u8d28\uff0c\u4f7f\u5176\u6700\u5c0f\u5316\u4e3a\u975e\u7279\u5b9a\u7684\u8fde\u63a5\u3002 Isso resulta em amostras de prote\u00ednas mais puras\u3002<\/li>\n<li><strong>\u4fbf\u5229\u6027\uff1a<\/strong> \u4f5c\u4e3a\u7b80\u5316\u7eaf\u5316\u8fc7\u7a0b\u7684\u78c1\u6027\u5c5e\u6027\u3002\u5982\u679c\u60a8\u60f3\u5feb\u901f\u5206\u79bb\uff0c\u53ef\u4ee5\u5feb\u901f\u5206\u79bb\u79bb\u5fc3\u6216\u590d\u5408\u8fc7\u6ee4\u6280\u672f\u3002<\/li>\n<li><strong>\u53ef\u6269\u5c55\u6027\uff1a<\/strong> \u534f\u8bae\u7684\u5927\u5c0f\u53d6\u51b3\u4e8e\u5b9e\u9a8c\u7684\u5fc5\u8981\u6027\uff0c\u4e0d\u540c\u7684\u76ee\u7684\u53ef\u80fd\u4f1a\u727a\u7272\u6548\u7387\u3002<\/li>\n<li><strong>\u65f6\u95f4\u6548\u7387\uff1a<\/strong> GFP Trap \u534f\u8bae\u5f88\u5feb\u5c31\u5b8c\u6210\u4e86\uff0c\u52a0\u901f\u4e86\u86cb\u767d\u8d28\u7eaf\u5316\u8fc7\u7a0b\uff0c\u5e76\u5141\u8bb8\u7814\u7a76\u4eba\u5458\u96c6\u4e2d\u5728\u540e\u7eed\u5e94\u7528\u4e2d\u3002<\/li>\n<\/ul>\n<h3>\u534f\u8bae\u6982\u8ff0<\/h3>\n<p>\u78c1\u6027 GFP \u9677\u9631\u534f\u8bae\u6d89\u53ca\u4e0d\u540c\u7684\u539f\u5219\uff1a<\/p>\n<ol>\n<li><strong>\u51c6\u5907\uff1a<\/strong> \u8bf7\u51c6\u5907\u597d\u7ec6\u80de\u4e0e GFP \u86cb\u767d\u7684\u7ade\u4e89\u3002\u79bb\u5fc3\u53bb\u9664\u7ec6\u80de\u788e\u5c51\u7684\u6f84\u6e05\u8bc1\u660e\u3002<\/li>\n<li><strong>\u8054\u8d5b:<\/strong> Adicione \u4f5c\u4e3a Esferas Magn\u00e9ticas GFP \u9677\u9631\uff0c\u4ee5\u6f84\u6e05\u548c\u5b75\u5316\u7684\u65b9\u5f0f\u6e29\u548c\u5730\u8868\u8fbe\u3002\u5141\u8bb8\u6355\u83b7 GFP \u86cb\u767d\u8d28\u3002<\/li>\n<li><strong>\u6d17\u6db2\uff1a<\/strong> \u4f5c\u4e3a\u4e00\u79cd\u8fde\u63a5\uff0c\u8bf7\u4f7f\u7528\u591a\u79cd\u65b9\u6cd5\u53bb\u9664\u86cb\u767d\u8d28\u548c\u6c61\u67d3\u7269\uff0c\u4ee5\u51c0\u5316\u86cb\u767d\u8d28\u3002<\/li>\n<li><strong>\u5220\u9664:<\/strong> \u6700\u540e\uff0c\u5c06\u86cb\u767d\u8d28\u4f5c\u4e3a\u86cb\u767d\u8d28\u7684\u6355\u83b7\u7269\uff0c\u4f7f\u7528\u9002\u5f53\u7684\u65b9\u6cd5\u6216\u5c06\u5176\u81ea\u7136\u5316\u4e3a\u86cb\u767d\u8d28\uff0c\u81ea\u7531\u5730\u4f5c\u4e3a afinidade \u7684\u76f8\u4e92\u4f5c\u7528\u3002<\/li>\n<\/ol>\n<h3>\u7ed3\u8bba<\/h3>\n<p>Esferas Magn\u00e9ticas GFP Trap \u534f\u8bae\u91c7\u7528\u7b80\u5355\u7684\u65b9\u6cd5\u548c\u9ad8\u6548\u7684\u86cb\u767d\u8d28\u7eaf\u5316\u65b9\u6cd5\uff0c\u7ed3\u5408\u9009\u62e9\u6027\u3001\u4fbf\u6377\u6027\u548c\u901f\u5ea6\u3002\u5229\u7528 GFP \u7684\u86cb\u767d\u8d28\u7279\u6027\uff0c\u53ef\u4ee5\u4e86\u89e3\u591a\u79cd\u5e94\u7528\u6240\u9700\u7684\u9ad8\u7eaf\u5ea6\u3001\u5171\u540c\u7814\u7a76\u3001\u529f\u80fd\u548c\u5f00\u53d1\u3002\u4f5c\u4e3a\u5bf9\u86cb\u767d\u8d28\u51c6\u5907\u5de5\u4f5c\u7684\u8981\u6c42\uff0c\u8fdb\u4e00\u6b65\u63d0\u9ad8\u86cb\u767d\u8d28\u7684\u8d28\u91cf\uff0c\u8be5\u534f\u8bae\u4ee3\u8868\u4e86\u5bf9\u86cb\u767d\u8d28\u7684\u4fdd\u62a4\u3002<\/p>\n<h2>O que Saber Antes de Usar o Protocolo das Esferas Magn\u00e9ticas GFP Trap<\/h2>\n<p>GFP Trap \u534f\u8bae\u662f\u5229\u7528\u751f\u7269\u5b66\u5206\u5b50\u6280\u672f\u5bf9\u7eff\u8272\u8367\u5149\u86cb\u767d (GFP) \u4e2d\u7684\u86cb\u767d\u8d28\u8fdb\u884c\u5206\u79bb\u548c\u7eaf\u5316\u3002\u91c7\u53d6\u5176\u4ed6\u6709\u6548\u7684\u65b9\u6cd5\u6765\u7814\u7a76\u4ea4\u4e92\u3001\u52a8\u529b\u5b66\u548c\u86cb\u767d\u8d28\u529f\u80fd\uff0c\u5728\u8fdb\u884c\u5b9e\u9a8c\u4e4b\u524d\u8003\u8651\u91cd\u8981\u7684\u5b58\u5728\u53d8\u5316\u3002<\/p>\n<h3>1. \u7406\u89e3 Liga\u00e7\u00e3o \u673a\u5236<\/h3>\n<p>O GFP Trap \u5229\u7528 esferas magn\u00e9ticas revestidas com anticorpos anti-GFP\u3002\u4f7f\u7528 GFP \u86cb\u767d\u8fdb\u884c\u8fde\u63a5\uff0c\u5141\u8bb8\u7814\u7a76\u4eba\u5458\u5c06\u5176\u89c6\u4e3a\u590d\u5408\u7269\u3002\u91cd\u8981\u7684\u662f\u8fde\u63a5\u7684\u673a\u5236\uff0cpH\u503c\u3001\u6e29\u5ea6\u548c\u529b\u91cf\u90fd\u4f1a\u5f71\u54cd\u8fde\u63a5\u7684\u6548\u7387\u3002<\/p>\n<h3>2. \u51c6\u5907Suas Amostras com Cuidado<\/h3>\n<p>\u534f\u8bae\u5f00\u59cb\u524d\u7684\u51c6\u5907\u5de5\u4f5c\u3001\u8bc1\u4e66\u7684\u51c6\u5907\u5de5\u4f5c\u5df2\u51c6\u5907\u5c31\u7eea\u3002\u8fd9\u5305\u62ec\u51c6\u5907\u5de5\u4f5c\uff0c\u4ee5\u53ca\u86cb\u767d\u8d28\u7684\u5efa\u7acb\u548c\u529f\u80fd\u7684\u5f00\u53d1\u6761\u4ef6\u3002\u53bb\u9664\u5bf9\u86cb\u767d\u9176\u6709\u653b\u51fb\u6027\u6216\u6291\u5236\u4f5c\u7528\u7684\u53bb\u57a2\u5242\uff0c\u53ef\u5e72\u6270\u53cd\u4f53\u7684\u8fde\u63a5\u3002\u4e3a\u786e\u4fdd GFP \u4e0e\u86cb\u767d\u8d28\u7684\u5747\u5300\u66dd\u5149\uff0c\u8bf7\u8fdb\u884c\u5747\u8d28\u5316\u3002<\/p>\n<h3>3. \u4f18\u5316\u4f5c\u4e3a Liga\u00e7\u00e3o \u6761\u4ef6<\/h3>\n<p>\u6211\u4eec\u901a\u8fc7\u5b9e\u9a8c\u6765\u8c03\u6574\u6bd4\u8d5b\u7684\u8282\u594f\u548c\u6e29\u5ea6\u3002\u4e00\u822c\u60c5\u51b5\u4e0b\uff0c\u5728 4\u00b0C \u7684\u6e29\u5ea6\u4e0b\u4fdd\u6e29 1-2 \u5c0f\u65f6\u662f\u7406\u60f3\u7684\u591a\u79cd\u534f\u8bae\uff0c\u53ef\u6d4b\u8bd5\u4e0d\u540c\u7684\u6761\u4ef6\u548c\u5173\u952e\u7684\u7ed3\u679c\u3002\u76d1\u63a7\u777e\u4e38\u9884\u8d5b\u671f\u95f4\u7684\u8bad\u7ec3\u6548\u7387\uff0c\u4ee5\u8c03\u6574\u534f\u594f\u66f2\u7684\u901f\u5ea6\u3002<\/p>\n<h3>4. \u7279\u6b8a\u8054\u8d5b\u7684\u6ce8\u610f\u4e8b\u9879<\/h3>\n<p>\u4e00\u4e2a\u7279\u6b8a\u7684\u8fde\u63a5\u610f\u5473\u7740\u4e00\u4e2a\u7eaf\u7cb9\u7684\u548c\u611f\u5174\u8da3\u7684\u86cb\u767d\u8d28\u7684\u7ed3\u5408\u3002\u5982\u679c\u60a8\u60f3\u91cd\u65b0\u5f00\u59cb\u6bd4\u8d5b\uff0c\u60a8\u53ef\u4ee5\u5728\u6bd4\u8d5b\u5f00\u59cb\u524d\u5b8c\u6210\u6240\u6709\u6bd4\u8d5b\u3002\u4e00\u4e2a\u8f85\u52a9\u6e05\u6d17\u7684\u5de5\u5177\u53ef\u4ee5\u6d88\u9664\u975e\u7279\u5b9a\u7684\u4ea4\u4e92\uff0c\u5e76\u4e14\u65e0\u9700\u5904\u7406\u4efb\u4f55\u86cb\u767d\u8d28\u3002<\/p>\n<h3>5. \u5207\u5c14\u6258\u65af\u63a7\u5236\u5b66\u6821<\/h3>\n<p>\u4f7f\u7528 GFP \u9677\u9631\u534f\u8bae\uff0c\u5305\u62ec\u6709\u6548\u63a7\u5236\u7ed3\u679c\u7684\u5fc5\u8981\u6761\u4ef6\u3002\u8bf7\u8003\u8651\u4f7f\u7528 GFP \u6765\u4e86\u89e3\u57fa\u91d1\u8054\u76df\u7684\u6700\u65b0\u60c5\u51b5\u3002\u6b64\u5916\uff0c\u6211\u4eec\u8fd8\u901a\u8fc7\u4e0d\u540c\u7684\u86cb\u767d\u8d28\u96c6\u4e2d\u5b9e\u9a8c\u6765\u5b9e\u73b0 GFP \u7684\u5e73\u884c\u5b9e\u9a8c\uff0c\u4ee5\u5efa\u7acb\u7279\u5b9a\u7684\u8fde\u63a5\u3002<\/p>\n<h3>6. \u8003\u8651\u5206\u6790\u6280\u672f<\/h3>\n<p>\u8fd9\u662f\u4e00\u79cd\u72ec\u7acb\u7684\u6280\u672f\u5206\u6790\u65b9\u6cd5\uff0c\u53ef\u4ee5\u89e3\u91ca\u7ed3\u679c\u3002\u5e38\u7528\u65b9\u6cd5\u5305\u62ec\u86cb\u767d\u8d28\u5370\u8ff9\u3001\u8d28\u8c31\u5206\u6790\u548c\u8367\u5149\u663e\u5fae\u955c\u3002\u719f\u6089\u51c0\u5316\u6280\u672f\u548c\u751f\u7269\u76f8\u5173\u6280\u672f\u3002<\/p>\n<h3>7. Esteja Ciente das Limita\u00e7\u00f5es Potenciais<\/h3>\n<p>Embora \u4f5c\u4e3a Esferas Magn\u00e9ticas GFP Trap \u53ef\u4ee5\u7eaf\u5316\u86cb\u767d\u8d28\uff0c\u5b58\u5728\u9650\u5236\u3002 Certas prote\u00ednas podem n\u00e3o se expressar bem com marcas de GFP or podem ser propensas \u00e0 agrega\u00e7\u00e3o.\u53e6\u5916\uff0c\u5728\u7ffb\u8bd1\u8fc7\u7a0b\u4e2d\u9700\u8981\u5bf9\u7ffb\u8bd1\u8fdb\u884c\u4fee\u6539\uff0c\u5e76\u786e\u4fdd\u534f\u8bae\u5728\u9694\u79bb\u8fc7\u7a0b\u4e2d\u4fdd\u7559\u4fee\u6539\u5185\u5bb9\u3002<\/p>\n<p>\u8bf7\u52a1\u5fc5\u8003\u8651\u4ee5\u4e0b\u56e0\u7d20\uff0c\u4ee5\u4fbf\u6700\u5927\u9650\u5ea6\u5730\u63d0\u9ad8 GFP \u78c1\u6027\u9677\u9631\u534f\u8bae\u7684\u529f\u6548\u5e76\u786e\u4fdd\u5b9e\u9a8c\u7ed3\u679c\u7684\u5b89\u5168\u6027\u3002\u55ef\uff0c\u98de\u673a\u548c\u6267\u884c\u662f\u4e3a\u4e86\u6df1\u5165\u4e86\u89e3\u611f\u5174\u8da3\u7684\u86cb\u767d\u8d28\u3002<\/p>\n<h2>Guia Passo a Passo do Protocolo de Esferas Magn\u00e9ticas GFP Trap<\/h2>\n<p>O Protocolo de Esferas Magn\u00e9ticas GFP Trap \u662f\u5229\u7528\u751f\u7269\u5b66\u5206\u5b50\u5bf9\u5206\u79bb\u7684\u86cb\u767d\u8d28\u548c\u7eff\u8272\u8367\u5149\u86cb\u767d (GFP) \u8fdb\u884c\u7814\u7a76\u7684\u65b9\u6cd5\u3002\u8fd9\u662f\u7814\u7a76\u4eba\u5458\u5bf9\u7ec6\u80de\u5185\u86cb\u767d\u8d28\u7684\u4ea4\u4e92\u3001\u529f\u80fd\u548c\u672c\u5730\u5316\u5206\u6790\u7684\u57fa\u672c\u8981\u6c42\u3002\u4e0b\u4e00\u6b65\u662f\u6267\u884c\u6709\u6548\u534f\u8bae\u3002<\/p>\n<h3>\u5fc5\u9700\u54c1<\/h3>\n<ul>\n<li>Esferas magn\u00e9ticas GFP Trap<\/li>\n<li>Lisato \u7ec6\u80de\u7ade\u4e89\u86cb\u767d marcadas com GFP<\/li>\n<li>\u8fde\u63a5\u7f13\u51b2\u6db2\uff08\u4f8b\u5982 PBS com \u9664\u57a2\u5242\uff09<\/li>\n<li>Buffer de lavagem\/Elui\u00e7\u00e3o<\/li>\n<li>\u4f0a\u739b<\/li>\n<li>Tubos de microcentr\u00edfuga<\/li>\n<li>\u5b9e\u9a8c\u5ba4\u79fb\u6db2\u7ba1\u548c\u79fb\u6db2\u7ba1\u6865<\/li>\n<\/ul>\n<h3>\u6b65\u9aa4 1\uff1a\u51c6\u5907 Lisato Cellular<\/h3>\n<p>Comece lisando suas c\u00e9lulas para liberar as prote\u00ednas\u3002\u4f7f\u7528\u9002\u5f53\u7684\u7f13\u51b2\u6db2\u53ef\u4ee5\u4fdd\u7559\u86cb\u767d\u8d28\u7684\u5b8c\u6574\u6027\uff0c\u4ece\u800c\u5141\u8bb8\u989d\u5916\u7684\u86cb\u767d\u8d28\u6807\u8bb0\u4e3a GFP\u3002\u5c06\u7ec6\u80de\u4f5c\u4e3a\u51dd\u80f6\u5b75\u80b2 30 \u5206\u949f\uff0c\u786e\u4fdd\u7ec6\u80de\u6210\u5206\u5145\u8db3\u3002 Uma vez feito\uff0c\u7528\u9ad8\u901f\u79bb\u5fc3\u673a\uff0812.000 g \u65cb\u8f6c\u529b\uff09\u5728 4\u00b0C \u4e0b\u53bb\u9664\u788e\u5c51 10-15 \u5206\u949f\u3002 Coleta o sobrenadante que cont\u00e9m suas prote\u00ednas marcadas com GFP\u3002<\/p>\n<h3>\u6b65\u9aa4 2\uff1a\u4f5c\u4e3a Esferas Magn\u00e9ticas GFP Trap \u7684\u8865\u5145<\/h3>\n<p>Adicione \u4f5c\u4e3a\u78c1\u529b GFP Trap \u6765\u6f84\u6e05\u3002\u6309\u6bd4\u4f8b\uff0c\u6bcf 10 \u00b5L \u7684\u5242\u91cf\u4e3a 1 mg \u7684\u86cb\u767d\u8d28\u603b\u91cf\uff0c\u53ef\u6839\u636e\u5177\u4f53\u9700\u8981\u8fdb\u884c\u7cbe\u786e\u8c03\u6574\u3002\u5728 30 \u5206\u949f\u7684\u73af\u5883\u6e29\u5ea6\u4e0b\uff0c\u5728\u5e73\u53f0\u4e0a\u65cb\u8f6c\u6216\u8f7b\u8f7b\u5730\u6405\u62cc\u3002\u6b64\u8fc7\u7a0b\u5141\u8bb8 GFP \u86cb\u767d\u4e0e\u78c1\u6027\u7269\u8d28\u53d1\u6325\u4f5c\u7528\u3002<\/p>\n<h3>\u7b2c\u4e09\u9636\u6bb5\uff1a\u62c9\u74e6\u5c14\u626e\u6f14\u57c3\u65af\u8d39\u62c9\u65af<\/h3>\n<p>Ap\u00f3s a incuba\u00e7\u00e3o, coloque o tube em um separador magn\u00e9tico para coletar as esferas rapidamente.\u79fb\u9664\u6216\u6270\u4e71\u9897\u7c92\u3002\u4f7f\u7528\u4e0d\u540c\u7684\u6e05\u6d17\u7f13\u51b2\u6db2\uff08\u6700\u591a 3-5 \u6b21\uff09\uff0c\u4ee5\u53bb\u9664\u86cb\u767d\u8d28\u7684\u7f13\u51b2\u6db2\u4f5c\u4e3a\u975e\u7279\u5b9a\u5f62\u5f0f\u7684\u6e05\u6d17\u5242\u3002\u83b7\u5f97\u5feb\u901f\u548c\u5b8c\u6574\u7684\u6d17\u6f31\u8bc1\u660e\uff0c\u5e76\u5728\u4f7f\u7528\u524d\u4e0e\u5176\u4ed6\u4eba\u5206\u5f00\u4f7f\u7528\u3002<\/p>\n<h3>\u6b65\u9aa4 4\uff1a\u827e\u9c81\u5c14 \u9970\u6f14 Prote\u00ednas Ligadas<\/h3>\n<p>\u4e3a\u4e86\u6062\u590d\u86cb\u767d\u8d28\uff0c\u8bf7\u4f7f\u7528 GFP \u78c1\u529b\uff0c\u5e76\u4f7f\u7528\u7f13\u51b2\u6db2\u3002\u5728\u73af\u5883\u6e29\u5ea6\u4e0b\u6d78\u6ce1\u7ea6 5-10 \u5206\u949f\uff0c\u4ee5\u4fdd\u8bc1\u86cb\u767d\u8d28\u7684\u5b89\u5168\u3002\u5728\u6b64\u8fc7\u7a0b\u4e2d\uff0c\u65b0\u7684\u7ba1\u5b50\u4e0e GFP \u7684\u86cb\u767d\u8d28\u5206\u79bb\u3002<\/p>\n<h3>\u6b65\u9aa4 5\uff1aAnalisar \u9970\u6f14 Prote\u00ednas Isoladas<\/h3>\n<p>\u6700\u540e\uff0c\u5bf9\u5206\u79bb\u7684\u86cb\u767d\u8d28\u8fdb\u884c\u5206\u6790\uff0c\u5e76\u4f7f\u7528 SDS-PAGE \u6280\u672f\u6216\u86cb\u767d\u8d28\u5370\u8ff9\u6cd5\u6765\u786e\u8ba4\u86cb\u767d\u8d28\u7684\u5b58\u5728\u548c\u7eaf\u5ea6\u3002\u6839\u636e\u5b9e\u9a8c\u6027\u8d28\uff0c\u6211\u4eec\u53ef\u4ee5\u6839\u636e\u9700\u8981\u8fdb\u884c\u529f\u80fd\u3001\u8d28\u91cf\u6216\u7279\u5f81\u5f62\u5f0f\u7684\u5149\u8c31\u6d4b\u91cf\u3002<\/p>\n<p>\u63a5\u4e0b\u6765\uff0c\u60a8\u53ef\u4ee5\u5229\u7528 GFP \u534f\u8bae\u7684\u6709\u6548\u529f\u80fd\uff0c\u5c06 GFP \u6355\u83b7\u5230\u5b64\u7acb\u7684\u86cb\u767d\u8d28\u4e2d\uff0c\u4ee5\u65b9\u4fbf\u540e\u7eed\u7684\u5206\u6790\u3002<\/p>\n<h2>Dicas Chave \u6267\u884c P\u00e9rolas Magn\u00e9ticas GFP Trap \u534f\u8bae\u7684 Bem-Sucedida<\/h2>\n<p>\u78c1\u77f3\u534f\u8bae\u7684\u5b9e\u65bd\u9700\u8981 GFP Trap \u7684\u8be6\u7ec6\u4fe1\u606f\u548c\u5b9e\u7528\u6280\u5de7\u3002\u6267\u884c\u5145\u5206\u7684\u64cd\u4f5c\u53ef\u7cbe\u786e\u83b7\u5f97 GFP\uff08Prote\u00edna Verde Fluorescente\uff09\u86cb\u767d\u8d28\u7684\u5206\u79bb\u548c\u5206\u6790\u7ed3\u679c\u3002\u8fd9\u662f\u4fdd\u8bc1\u5de5\u4f5c\u987a\u5229\u548c\u6709\u6548\u7684\u5173\u952e\u3002<\/p>\n<h3>1. Otimize \u4f5c\u4e3a Condi\u00e7\u00f5es de Lise Cellular<\/h3>\n<p>\u7ec6\u80de\u7684\u6709\u6548\u5229\u7528\u5bf9\u4e8e GFP \u86cb\u767d\u8d28\u7684\u91ca\u653e\u81f3\u5173\u91cd\u8981\u3002\u4f7f\u7528\u6e05\u6d01\u5242\u548c\u9002\u5f53\u7684\u6e05\u6d01\u5242\u6765\u6eb6\u89e3\u819c\u3002\u6211\u5df2\u7ecf\u4e86\u89e3\u4e86\u777e\u4e38\u7684\u521d\u6b65\u60c5\u51b5\uff0c\u4ee5\u51b3\u5b9a GFP \u7684\u6700\u5927\u5316\u6216\u86cb\u767d\u8d28\u964d\u89e3\u7684\u901f\u5ea6\u548c\u6e29\u5ea6\u3002<\/p>\n<h3>2. \u4f7f\u7528 P\u00e9rolas Magn\u00e9ticas Preparadas Recentemente<\/h3>\n<p>\u7531\u4e8e\u78c1\u6027\u78c1\u77f3\u5728\u751f\u547d\u6709\u9650\u7684\u60c5\u51b5\u4e0b\uff0c\u6548\u7387\u4f1a\u964d\u4f4e\u3002\u59cb\u7ec8\u786e\u4fdd\u60a8\u53ef\u4ee5\u4f7f\u7528\u6700\u8fd1\u51c6\u5907\u7684\u8bba\u575b\u6216\u5728\u5236\u9020\u8fc7\u7a0b\u4e2d\u4f7f\u7528 acordo \u8bbe\u5907\u3002\u5982\u679c\u6ca1\u6709\u8db3\u591f\u7684\u80fd\u529b\u6765\u5b8c\u6210\u8054\u8d5b\uff0c\u5c31\u4f1a\u5f71\u54cd\u5230\u534f\u8bae\u7684\u6210\u529f\u3002<\/p>\n<h3>3. Mantenha a Concentra\u00e7\u00e3o Adequada da Amostra<\/h3>\n<p>\u7ec6\u80de\u6216\u86cb\u767d\u8d28\u7684\u6d53\u7f29\u7269\uff0c\u53ef\u4ee5\u5728\u4e0d\u540c\u65f6\u95f4\u95f4\u9694\u5185\u4ea7\u751f\u3002\u96c6\u4e2d\u7cbe\u529b\u5c06\u73cd\u73e0\u9971\u548c\uff0c\u5e76\u4e14\u5c06\u5176\u5145\u5206\u5229\u7528\u3002\u5b9e\u73b0\u51b3\u5b9a\u6027\u7684\u5b9e\u9a8c\uff0c\u4f5c\u4e3a\u83b7\u5f97\u6700\u4f73\u7ed3\u679c\u7684\u7279\u5b9a\u60f3\u6cd5\u7684\u6761\u4ef6\u3002<\/p>\n<h3>4. \u786e\u4fdd\u6df7\u5408\u7269\u5b8c\u5168<\/h3>\n<p>\u4f5c\u4e3a\u8fde\u63a5\u548c\u704c\u6d17\u7684\u6301\u7eed\u65f6\u95f4\uff0c\u5b83\u662f\u4e0e GFP \u548c\u73cd\u73e0\u5ca9\u7b49\u86cb\u767d\u8d28\u76f8\u4e92\u4f5c\u7528\u6700\u5927\u5316\u7684\u5fc5\u8981\u8865\u5145\u3002\u4f7f\u7528\u6e29\u548c\u7684\u7ba1\u9053\u6216\u65cb\u8f6c\u88c5\u7f6e\u6765\u6355\u83b7\u86cb\u767d\u8d28\u3002<\/p>\n<h3>5. \u4f7f\u7528\u9002\u5f53\u7684\u6d17\u6db2<\/h3>\n<p>\u704c\u6d17\u7684\u5173\u952e\u7ec4\u4ef6\u662f\u9488\u5bf9\u975e\u7279\u5b9a\u60c5\u51b5\u7684\u53bb\u9664\u86cb\u767d\u7684\u534f\u8bae\u3002\u786e\u4fdd\u5b9e\u73b0\u704c\u6d17\u7684\u5404\u79cd\u53d8\u5316\uff0c\u4ee5\u9632\u6b62\u5bf9\u7279\u5b9a\u7684\u4f69\u7f57\u62c9\u548c\u86cb\u767d\u8d28\u7684\u5f71\u54cd\u3002\u5982\u679c\u6ca1\u6709\uff0c\u8bf7\u4e0d\u8981\u6309\u538b\u6216\u6309\u538b\u704c\u6d17\u7a0b\u5e8f\uff0c\u5426\u5219\u6700\u7ec8\u4f1a\u9020\u6210\u6c61\u67d3\u3002<\/p>\n<h3>6. \u4f18\u5316\u4f5c\u4e3a Elui\u00e7\u00e3o \u6761\u4ef6<\/h3>\n<p>\u8fdb\u884c\u8fde\u63a5\u548c\u704c\u6d17\uff0c\u4ee5\u63d0\u53d6\u78c1\u77f3\u4e2d\u7684\u86cb\u767d\u8d28\u3002\u4f7f\u7528 GFP \u6765\u91ca\u653e\u86cb\u767d\u8d28\u7684\u6761\u4ef6\u3002\u53ef\u4ee5\u8f7b\u677e\u5730\u8fdb\u884c\u5b9e\u9a8c\uff0c\u5305\u62ec\u4e0d\u540c\u7684 pH \u503c\u6216\u7ade\u4e89\u6027\u8bd5\u5242\uff0c\u4ee5\u53ca\u5176\u4ed6\u670d\u52a1\u3002\u6d4b\u8bd5\u5404\u79cd\u786e\u5b9a\u7279\u5b9a\u86cb\u767d\u8d28\u7684\u6709\u6548\u534f\u8bae\u7684\u8ba1\u7b97\u901f\u5ea6\u3002<\/p>\n<h3>7. \u9a8c\u8bc1\u7ed3\u679c<\/h3>\n<p>\u4f5c\u4e3a\u534f\u8bae\u7684\u4e00\u90e8\u5206\uff0c\u5b83\u662f GFP \u7ed3\u6784\u4e2d\u86cb\u767d\u8d28\u9694\u79bb\u7684\u91cd\u8981\u9a8c\u8bc1\u3002\u6280\u672f\u5305\u62ec SDS-PAGE\u3001\u86cb\u767d\u8d28\u5370\u8ff9\u3001\u8367\u5149\u663e\u5fae\u955c\u7b49\uff0c\u4ee5\u786e\u8ba4\u5206\u79bb\u3002\u6267\u884c\u5668\u63a7\u5236\u6240\u6709\u64cd\u4f5c\u5e76\u786e\u4fdd\u673a\u5bc6\u548c\u518d\u73b0\u7684\u7ed3\u679c\u3002<\/p>\n<h3>8. \u6ce8\u518c Tudo<\/h3>\n<p>\u8be6\u7ec6\u7684\u8be6\u7ec6\u8fc7\u7a0b\u6587\u6863\u5141\u8bb8\u91cd\u590d\u548c\u89e3\u51b3\u95ee\u9898\u3002\u6ce8\u518c\u8be6\u7ec6\u4fe1\u606f\u3001\u5b75\u5316\u901f\u5ea6\u548c\u6e29\u5ea6\u7684\u7ec4\u5408\u3002\u91cd\u8981\u7684\u662f\u8981\u91cd\u73b0\u7ed3\u679c\uff0c\u5e76\u8bc6\u522b\u672a\u6765\u5b9e\u9a8c\u4e2d\u6240\u5fc5\u9700\u7684\u7f8e\u597d\u7ed3\u679c\u3002<\/p>\n<p>\u4ece\u6839\u672c\u4e0a\u8bb2\uff0c\u6211\u4eec\u53ef\u4ee5\u6210\u529f\u5730\u5b8c\u6210\u78c1\u6027 GFP \u9677\u9631\u534f\u8bae\uff0c\u5e76\u5728\u86cb\u767d\u8d28\u5206\u79bb\u5b9e\u9a8c\u4e2d\u83b7\u5f97\u7ed3\u679c\u548c\u91cd\u73b0\u3002<\/p>","protected":false},"excerpt":{"rendered":"<p>No campo da biologia molecular, o Protocolo de Beads Magn\u00e9ticas GFP Trap tornou-se uma t\u00e9cnica fundamental para isolar e purificar prote\u00ednas marcadas com Prote\u00edna Fluorescente Verde (GFP). Este m\u00e9todo inovador melhora a purifica\u00e7\u00e3o de prote\u00ednas aproveitando as propriedades de liga\u00e7\u00e3o \u00fanicas de beads magn\u00e9ticas revestidas com anticorpos que visam especificamente a GFP. A efic\u00e1cia deste [&hellip;]<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"site-sidebar-layout":"default","site-content-layout":"","ast-site-content-layout":"default","site-content-style":"default","site-sidebar-style":"default","ast-global-header-display":"","ast-banner-title-visibility":"","ast-main-header-display":"","ast-hfb-above-header-display":"","ast-hfb-below-header-display":"","ast-hfb-mobile-header-display":"","site-post-title":"","ast-breadcrumbs-content":"","ast-featured-img":"","footer-sml-layout":"","ast-disable-related-posts":"","theme-transparent-header-meta":"","adv-header-id-meta":"","stick-header-meta":"","header-above-stick-meta":"","header-main-stick-meta":"","header-below-stick-meta":"","astra-migrate-meta-layouts":"default","ast-page-background-enabled":"default","ast-page-background-meta":{"desktop":{"background-color":"","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"tablet":{"background-color":"","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"mobile":{"background-color":"","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""}},"ast-content-background-meta":{"desktop":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"tablet":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"mobile":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""}},"footnotes":""},"categories":[1],"tags":[],"class_list":["post-7975","post","type-post","status-publish","format-standard","hentry","category-news"],"_links":{"self":[{"href":"https:\/\/nanomicronspheres.com\/zh\/wp-json\/wp\/v2\/posts\/7975","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/nanomicronspheres.com\/zh\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/nanomicronspheres.com\/zh\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/nanomicronspheres.com\/zh\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/nanomicronspheres.com\/zh\/wp-json\/wp\/v2\/comments?post=7975"}],"version-history":[{"count":0,"href":"https:\/\/nanomicronspheres.com\/zh\/wp-json\/wp\/v2\/posts\/7975\/revisions"}],"wp:attachment":[{"href":"https:\/\/nanomicronspheres.com\/zh\/wp-json\/wp\/v2\/media?parent=7975"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/nanomicronspheres.com\/zh\/wp-json\/wp\/v2\/categories?post=7975"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/nanomicronspheres.com\/zh\/wp-json\/wp\/v2\/tags?post=7975"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}