{"id":252,"date":"2025-01-26T22:44:56","date_gmt":"2025-01-26T14:44:56","guid":{"rendered":"https:\/\/tiand.top\/?p=252"},"modified":"2025-01-28T09:16:11","modified_gmt":"2025-01-28T01:16:11","slug":"%e5%b0%8f%e9%bc%a0%e8%82%bf%e7%98%a4%e7%bb%84%e7%bb%87%e8%a7%a3%e7%a6%bb%e5%92%8c%e7%bb%86%e8%83%9e%e5%88%86%e7%a6%bb","status":"publish","type":"post","link":"https:\/\/tiand.top\/index.php\/2025\/01\/26\/%e5%b0%8f%e9%bc%a0%e8%82%bf%e7%98%a4%e7%bb%84%e7%bb%87%e8%a7%a3%e7%a6%bb%e5%92%8c%e7%bb%86%e8%83%9e%e5%88%86%e7%a6%bb\/","title":{"rendered":"\u5c0f\u9f20\u80bf\u7624\u7ec4\u7ec7\u89e3\u79bb\u548c\u7ec6\u80de\u5206\u79bb"},"content":{"rendered":"\n

\u3010\u5b9e\u9a8c\u8bd5\u5242\u53ca\u6750\u6599\u3011<\/strong><\/h4>\n\n\n\n
    \n
  1. MACS buffer\uff1a500 mL DPBS + 2.5 g BSA \uff08Sigma, Cat: B2064\uff09+ 2 mL 0.5M EDTA\uff08Thermo, Cat: AM9260G\uff09<\/li>\n\n\n\n
  2. \u8367\u5149\u6807\u8bb0\u7684\u7ec6\u80de\u8868\u9762\u5206\u5b50\u76f8\u5173\u6297\u4f53<\/li>\n\n\n\n
  3. 5 mL \u6d41\u5f0f\u7ba1\uff08BD Falcon\uff09<\/li>\n\n\n\n
  4. U\u5e9596\u5b54\u677f1 mL\u6ce8\u5c04\u5668<\/li>\n\n\n\n
  5. 10 mL\u6ce8\u5c04\u566870 um\u7b5b\u7f516\u5b54\u7ec6\u80de\u57f9\u517b\u677f<\/li>\n\n\n\n
  6. 50 mL\u79bb\u5fc3\u7ba1\uff0c15 mL\u79bb\u5fc3\u7ba1<\/li>\n\n\n\n
  7. 30% Percoll\uff08GE, Cat: 17089109\uff09\uff1a1\u4efd 10X HBSS + 9\u4efd Percoll\u539f\u6db2\u2192100% Percoll\u2192+1X HBSS \u7a00\u91ca\u81f330%\uff0830%\u5305\u542b\u9ad3\u7cfb\u7ec6\u80de\uff0c\u53ea\u7814\u7a76\u6dcb\u5df4\u7ec6\u80de\u53ef\u752840%\uff09<\/li>\n\n\n\n
  8. \u6d88\u5316\u6db2\u914d\u5236\u30106ml\/\u4e2a\u80bf\u7624\uff08\u5355\u4e2a\u6700\u59271.5cm3<\/sup>\uff09\u3011\uff1a1:50\uff08120ul\uff09 RPMI1640+10%\u80ce\u725b\u8840\u6e05+1mg\/mL\u80f6\u539f\u86cb\u767d\u9176IV+0.1mg\/mL DNase\uff0c1640\u7a00\u91ca\u81f36mL<\/li>\n\n\n\n
  9. \u80f6\u539f\u86cb\u767d\u9176IV\uff08Worthington\uff0cCat: LS004189\uff09\u50a8\u5907\u6db2\uff1aHBSS\u7a00\u91ca\u81f350mg\/ml<\/li>\n\n\n\n
  10. DNase\uff08Sigma\uff0cCat: DN25\uff09\u50a8\u5907\u6db2\uff1a1640\u7a00\u91ca\u81f350mg\/ml<\/li>\n\n\n\n
  11. \u56fd\u4ea7\u80ce\u725b\u8840\u6e05\uff08\u6d88\u5316\u80bf\u7624\u7528\uff0c\u666e\u8bfa\u8d5b\uff0cC<\/a>at: 164210\uff09<\/li>\n\n\n\n
  12. \u7ea2\u7ec6\u80de\u88c2\u89e3\u6db2\uff08\u78a7\u4e91\u5929\uff0cCat: C3702\uff09<\/li>\n<\/ol>\n\n\n\n

    \u3010\u5b9e\u9a8c\u5185\u5bb9\u3011<\/strong><\/h4>\n\n\n\n
      \n
    1. \u53d6\u8377\u7624\u5c0f\u9f20\u80bf\u7624\u7ec4\u7ec7\uff0c\u6d78\u6ce1\u5728\u542b1%\u80ce\u725b\u8840\u6e05+1%\u9752\u94fe\u9709\u7d20\u7684PBS\u4e2d\uff0c\u572810cm\u57f9\u517b\u76bf\u4e2d\u7528\u65e0\u83cc\u7684\u773c\u79d1\u526a\u5c06\u80bf\u7624\u7ec4\u7ec7\u526a\u788e\u81f3\u7ea62-3mm3<\/sup>\u3002<\/li>\n\n\n\n
    2. 500g\u00d75min\u79bb\u5fc3\u3002<\/li>\n\n\n\n
    3. \u5f03\u4e0a\u6e05\uff0c\u7528\u80bf\u7624\u7ec4\u7ec7\u6d88\u5316\u6db2\u91cd\u60ac\u526a\u788e\u7684\u80bf\u7624\u7ec4\u7ec7\uff0c37\u2103\u6052\u6e29\u6447\u5e8a160rpm  2\u5c0f\u65f6\u3002\u80bf\u7624\u7ec4\u7ec7\u6d88\u5316\u6db2\u4e3a\uff1aRPMI1640<\/a>+10%\u80ce\u725b\u8840\u6e05+1mg\/mL\u80f6\u539f\u86cb\u767d\u9176IV+0.1mg\/mL DNase\u3002\uff08\u6d88\u5316\u6db26ml\/\u4e2a\u80bf\u7624\uff09\u3002<\/li>\n\n\n\n
    4. \u5c06\u7ec4\u7ec7\u5012\u516510cm\u57f9\u517b\u76bf\u4e2d\uff0c\u52a0\u5165PBS\u7ec8\u6b62\uff0c\u752810mL\u6ce8\u5c04\u5668\u53cd\u590d\u5439\u6253\u6d88\u5316\u540e\u7684\u80bf\u7624\u7ec4\u7ec7\u540e\uff0c\u752870\u03bcm\u6ee4\u7f51\u8fc7\u6ee4\u3002<\/li>\n\n\n\n
    5. \u6536\u96c6\u6ee4\u6db2\u540e500g\u00d7<\/a>5min\u79bb\u5fc3\uff0c\u7528\u7ea2\u7ec6\u80de\u88c2\u89e3\u6db23-4ml\/\u7ba1\u91cd\u60ac\u7ec6\u80de\u6c89\u6dc0\uff0c\u51b0\u4e0a\u88c2\u89e35min\u3002<\/li>\n\n\n\n
    6. \u52a0\u51652\u500d\u4f53\u79ef\u7684PBS\u7ec8\u6b62\u53cd\u5e94\uff0c500g\u00d75min<\/a>\u79bb\u5fc3\u3002<\/li>\n\n\n\n
    7. \u8fc730% Percoll\uff1a\u6bcf\u7ba1\u52a0\u516512 ml 30% Percoll\u91cd\u60ac\u6c89\u6dc0\uff0c\u5ba4\u6e291000g\u00d730min\uff0c\u5347\u4e00\u964d\u4e00\uff0c45min\u3002<\/li>\n\n\n\n
    8. \u5c0f\u5fc3\u5438\u8d70\u4e0a\u6e05\uff0c\u52a0\u5165PBS\u6d17\u4e00\u6b21\uff0c500g\u00d75min\u79bb\u5fc3\u540e\uff0c\u52a0\u5165PBS\u91cd\u60ac\u6c89\u6dc0\uff0c\u5c06\u7ec6\u80de\u60ac\u6db2\u79fb\u5230\u6d41\u5f0f\u7ba1\u4e2d\u5f85\u6d41\u5f0f\u67d3\u8272\u3002<\/li>\n<\/ol>\n","protected":false},"excerpt":{"rendered":"

      \u3010\u5b9e\u9a8c\u8bd5\u5242\u53ca\u6750\u6599\u3011 \u3010\u5b9e\u9a8c\u5185\u5bb9\u3011<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_jetpack_memberships_contains_paid_content":false,"footnotes":""},"categories":[8],"tags":[],"class_list":["post-252","post","type-post","status-publish","format-standard","hentry","category-protocols"],"jetpack_featured_media_url":"","jetpack_sharing_enabled":true,"_links":{"self":[{"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/posts\/252","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/comments?post=252"}],"version-history":[{"count":2,"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/posts\/252\/revisions"}],"predecessor-version":[{"id":296,"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/posts\/252\/revisions\/296"}],"wp:attachment":[{"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/media?parent=252"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/categories?post=252"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/tiand.top\/index.php\/wp-json\/wp\/v2\/tags?post=252"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}