{"id":1738,"date":"2017-07-16T22:48:21","date_gmt":"2017-07-16T14:48:21","guid":{"rendered":"http:\/\/chundubio.com\/?p=1738"},"modified":"2017-07-16T22:48:21","modified_gmt":"2017-07-16T14:48:21","slug":"%e5%8d%95%e5%85%8b%e9%9a%86%e6%8a%97%e4%bd%93%e5%8d%95%e6%8a%97%e7%9a%84%e7%ba%af%e5%8c%96%e6%96%b9%e6%b3%95","status":"publish","type":"post","link":"http:\/\/chundubio.com\/?p=1738","title":{"rendered":"\u5355\u514b\u9686\u6297\u4f53\/\u5355\u6297\u7684\u7eaf\u5316\u65b9\u6cd5"},"content":{"rendered":"
\u4e00\u3001\u8179\u6c34\u578b\u5355\u6297\u7684\u7eaf\u5316<\/b><\/div>\n
\u5728\u5355\u6297\u7eaf\u5316\u4e4b\u524d\uff0c\u4e00\u822c\u5747\u9700\u5bf9\u8179\u6c34\u8fdb\u884c\u9884\u5904\u7406\uff0c\u76ee\u7684\u662f\u4e3a\u4e86\u8fdb\u4e00\u6b65\u9664\u53bb\u7ec6\u80de\u53ca\u5176\u6b8b\u6e23\u3001\u5c0f\u9897\u7c92\u7269\u8d28\u3001\u4ee5\u53ca\u8102\u80aa\u6ef4\u7b49\u3002\u5e38\u7528\u7684\u65b9\u6cd5\u6709\u4e8c\u6c27\u5316\u7845\u5438\u9644\u6cd5\u548c\u8fc7\u6ee4\u79bb\u5fc3\u6cd5\uff0c\u4ee5\u524d\u8005\u5904\u7406\u6548\u679c\u4e3a\u4f73\uff0c\u800c\u4e14\u64cd\u4f5c\u7b80\u4fbf\u3002<\/div>\n
1<\/b>\u3001\u4e8c\u6c27\u5316\u7845\u5438\u9644\u6cd5<\/b><\/div>\n
\u65b0\u9c9c\u91c7\u96c6\u7684\u8179\u6c34\uff08\u6216\u51bb\u5b58\u7684\u8179\u6c34\uff09\uff0c2000r\/min 15\u5206\u949f\uff0c\u9664\u53bb\u7ec6\u80de\u6210\u5206\uff08\u6216\u51bb\u5b58\u8fc7\u7a0b\u4e2d\u5f62\u6210\u7684\u56fa\u4f53\u7269\u8d28\uff09\u7b49\uff1b\u53d6\u4e0a\u5c42\u6e05\u4eae\u7684\u8179\u6c34\uff0c\u7b49\u91cf\u52a0\u5165PH7.2\u5df4\u6bd4\u59a5\u7f13\u51b2\u76d0\u6c34\uff08VBS\uff1b0.004mol\/L\u5df4\u6bd4\u59a5\uff0c0.15mol\/L NaCl\uff0c0.8mmol\/L Mg2+<\/sup>\uff0c0.3mmol\/L Ca2+<\/sup>\uff09\u7a00\u91ca\uff1b\u7136\u540e\u4ee5\u6bcf10ml\u7a00\u91ca\u8179\u6c34\u4e2d\u52a0150mg\u4e8c\u6c27\u5316\u7845\u7c89\u672b\uff0c\u6df7\u5300\uff0c\u60ac\u6db2\u5728\u5ba4\u6e29\u5b75\u80b230\u5206\u949f\uff0c\u4e0d\u65f6\u6447\u52a8\uff1b2000g\u79bb\u5fc320\u5206\u949f\uff0c\u8102\u8d28\u7b49\u901a\u8fc7\u8be5\u6cd5\u9664\u53bb\uff0c\u5373\u53ef\u5f97\u6f84\u6e05\u7684\u8179\u6c34\u3002<\/div>\n
2<\/b>\u3001\u8fc7\u6ee4\u79bb\u5fc3\u6cd5<\/b><\/div>\n
\u7528\u5fae\u5b54\u6ee4\u819c\u8fc7\u6ee4\u8179\u6c34\uff0c\u4ee5\u9664\u53bb\u8f83\u5927\u7684\u51dd\u5757\u53ca\u8102\u80aa\u6ef4\uff1b\u752810000g 15\u5206\u949f\u9ad8\u901f\u79bb\u5fc3\uff084\u2103\uff09\u9664\u53bb\u7ec6\u80de\u6b8b\u6e23\u53ca\u5c0f\u9897\u7c92\u7269\u8d28\u3002<\/div>\n
3<\/b>\u3001\u6df7\u5408\u6cd5<\/b><\/div>\n
\u5373\u4e0a\u8ff0\u4e24\u6cd5\u7684\u7ec4\u5408\uff0c\u5148\u5c06\u8179\u6c34\u9ad8\u901f\u79bb\u5fc3\uff0c\u53d6\u4e0a\u6e05\u6db2\u518d\u7528\u4e8c\u6c27\u5316\u7845\u5438\u9644\u5904\u7406\u3002<\/div>\n
\u4e8c\u3001\u5355\u6297\u7684\u7c97\u63d0<\/b><\/div>\n
1<\/b>\u3001\u786b\u9178\u94f5\u6c89\u6dc0\u6cd5<\/b><\/div>\n
\uff081\uff09\u9971\u548c\u786b\u9178\u94f5\u6eb6\u6db2\u7684\u914d\u5236<\/div>\n
500g\u786b\u9178\u94f5\u52a0\u5165500ml\u84b8\u998f\u6c34\u4e2d\uff0c\u52a0\u70ed\u81f3\u5b8c\u5168\u6eb6\u89e3\uff0c\u5ba4\u6e29\u8fc7\u591c\uff0c\u6790\u51fa\u7684\u7ed3\u6676\u4efb\u5176\u7559\u5728\u74f6\u4e2d\u3002\u4e34\u7528\u524d\u53d6\u6240\u9700\u7684\u91cf\uff0c\u75282mol\/L NaOH\u8c03PH\u81f37.8\u3002<\/div>\n
\uff082\uff09\u76d0\u6790<\/div>\n
\u5438\u53d610ml\u5904\u7406\u597d\u7684\u8179\u6c34\u79fb\u5165\u5c0f\u70e7\u676f\u4e2d\uff0c\u5728\u6405\u62cc\u4e0b\uff0c\u6ef4\u52a0\u9971\u548c\u786b\u9178\u94f5\u6eb6\u6db25.0ml\uff1b\u7ee7\u7eed\u7f13\u6162\u6405\u62cc30\u5206\u949f\uff1b10000r\/min\u79bb\u5fc315\u5206\u949f\uff1b\u5f03\u53bb\u4e0a\u6e05\u6db2\uff0c\u6c89\u6dc0\u7269\u75281\/3\u9971\u548c\u5ea6\u786b\u9178\u94f5\u60ac\u6d6e\uff0c\u6405\u62cc\u4f5c\u752830\u5206\u949f\uff0c\u540c\u6cd5\u79bb\u5fc3\uff1b\u91cd\u590d\u524d\u4e00\u6b651-2\u6b21\uff1b\u6c89\u6dc0\u7269\u6eb6\u4e8e1.5ml PBS\uff080.01mol\/L PH7.2\uff09\u6216Tris-HCl\u7f13\u51b2\u6db2\u4e2d\u3002<\/div>\n
\uff083\uff09\u8131\u76d0<\/div>\n
\u5e38\u7528\u67f1\u5c42\u6790\u6216\u900f\u6790\u6cd5\u3002\u67f1\u5c42\u6790\u6cd5\u662f\u5c06\u76d0\u6790\u6837\u54c1\u8fc7Sephadex G-50\u5c42\u6790\u67f1\uff0c\u4ee5PBS\u6216Tris-HCl\u7f13\u51b2\u6db2\u4f5c\u4e3a\u5e73\u8861\u6db2\u548c\u6d17\u8131\u6db2\uff0c\u6d41\u901f\u6bcf\u5206\u949f1ml\u3002\u7b2c\u4e00\u4e2a\u86cb\u767d\u5cf0\u5373\u4e3a\u8131\u76d0\u7684\u6297\u4f53\u6eb6\u6db2\u3002\u900f\u6790\u6cd5\u662f\u5c06\u900f\u6790\u888b\u4e8e2% NaHCO3<\/sub>\uff0c1mmol\/LEDTA\u6eb6\u6db2\u4e2d\u716e10\u5206\u949f\uff0c\u7528\u84b8\u998f\u6c34\u6e05\u6d17\u900f\u6790\u888b\u5185\u5916\u8868\u9762\uff0c\u518d\u7528\u84b8\u998f\u6c34\u716e\u900f\u6790\u888b10\u5206\u949f\uff0c\u51b7\u81f3\u5ba4\u6e29\u5373\u53ef\u4f7f\u7528\uff08\u5e76\u53ef\u4e8e0.2mol\/L\u00a0<\/span>EDTA\u6eb6\u6db2\u4e2d\uff0c4\u2103\u4fdd\u5b58\u5907\u7528\uff09\u3002\u5c06\u76d0\u6790\u6837\u54c1\u88c5\u5165\u900f\u6790\u888b\u4e2d\uff0c\u5bf950-100\u500d\u4f53\u79ef\u7684PBS\u6216Tris-HCl\u7f13\u51b2\u6db2\u900f\u6790\uff084\u2103\uff0912-24\u5c0f\u65f6\uff0c\u5176\u95f4\u66f4\u6362 5\u6b21\u900f\u6790\u6db2\uff0c\u7528\u8418\u6c0f\u8bd5\u5242\uff08\u7898\u5316\u6c5e11.5g\uff0c\u7898\u5316\u94be8g\uff0c\u52a0\u84b8\u998f\u6c3450ml\uff0c\u5f85\u6eb6\u89e3\u540e\uff0c\u518d\u52a020% NaOH 50ml\uff09\u68c0\u6d4b\uff0c\u76f4\u81f3\u900f\u6790\u5916\u6db2\u65e0\u9ec4\u8272\u7269\u5f62\u6210\u4e3a\u6b62\u3002<\/div>\n
\uff084\uff09\u86cb\u767d\u8d28\u542b\u91cf\u7684\u6d4b\u5b9a<\/div>\n
\uff08Pr\uff09\uff08mg\/ml\uff09=\uff081.45\u00d7OD280-0.74\u00d7OD260\uff09\u00d7\u7a00\u91ca\u500d\u6570\uff1b\u6216\uff08Pr\uff09=OD280\u00d7\u7a00\u91ca\u500d\u6570\/1.3<\/div>\n
\uff085\uff09\u5206\u88c5\u51bb\u5b58\u5907\u7528<\/div>\n
2<\/b>\u3001\u8f9b\u9178<\/b>–<\/b>\u786b\u9178\u94f5\u6c89\u6dc0\u6cd5<\/b><\/div>\n
\u8be5\u6cd5\u7b80\u5355\u6613\u884c\uff0c\u9002\u5408\u4e8e\u63d0\u7eafIgG1\u548cIgG2b\uff0c\u4f46\u5bf9IgG3\u548cIgA\u7684\u56de\u6536\u7387\u53ca\u7eaf\u5316\u6548\u679c\u5dee\u3002\u5176\u4e3b\u8981\u6b65\u9aa4\u5982\u4e0b\uff1a\u53d61\u4efd\u9884\u5904\u7406\u8fc7\u7684\u8179\u6c34\u52a02\u4efd 0.06mol\/L PH5.0\u918b\u9178\u7f13\u51b2\u6db2\uff0c\u75281mol\/L HCl\u8c03PH\u81f34.8\uff1b\u6309\u6bcf\u6beb\u5347\u7a00\u91ca\u8179\u6c34\u52a011ul\u8f9b\u9178\u7684\u6bd4\u4f8b\uff0c\u5ba4\u6e29\u6405\u62cc\u4e0b\u9010\u6ef4\u52a0\u5165\u8f9b\u9178\uff0c\u4e8e30\u5206\u949f\u5185\u52a0\u5b8c\uff0c4\u2103\u9759\u7f6e2\u5c0f\u65f6\uff0c\u53d6\u51fa15000g\u79bb\u5fc330\u5206\u949f\uff0c\u5f03\u6c89\u6dc0\uff1b\u4e0a\u6e05\u7ecf\u5c3c\u9f99\u7b5b\u8fc7\u6ee4\uff08125um\uff09\uff0c\u52a0\u51651\/10\u4f53\u79ef\u76840.01mol\/L PBS\uff0c\u75281mol\/L NaOH\u8c03PH\u81f37.2\uff1b\u57284\u2103\u4e0b\u52a0\u5165\u9971\u548c\u786b\u9178\u94f5\u81f345%\u9971\u548c\u5ea6\uff0c\u4f5c\u752830\u5206\u949f\uff0c\u9759\u7f6e1\u5c0f\u65f6\uff1b10000g\u79bb\u5fc330\u5206\u949f\uff0c\u5f03\u4e0a\u6e05\uff1b\u6c89\u6dc0\u6eb6\u4e8e\u9002\u91cfPBS\uff08\u542b 137mmol\/L NaCl\uff0c2.6mol\/L KCl\uff0c0.2mmol\/L EDTA\uff09\u4e2d\uff0c\u5bf950-100\u500d\u4f53\u79ef\u7684PBS\u900f\u6790\uff0c4\u2103\u8fc7\u591c\uff0c\u5176\u95f4\u6362\u6c343\u6b21\u4ee5\u4e0a\uff1b\u53d6\u51fa10000g\u79bb\u5fc330\u5206\u949f\uff0c\u9664\u53bb\u4e0d\u6eb6\u6027\u6c89\u6e23\uff0c\u6d4b\u5b9a\u86cb\u767d\u8d28\u542b\u91cf\u540e\uff0c\u5206\u88c5\uff0c\u51bb\u5b58\u5907\u7528\u3002<\/div>\n
3<\/b>\u3001\u4f18\u7403\u86cb\u767d\u6c89\u6dc0\u6cd5<\/b><\/div>\n
\u8be5\u6cd5\u9002\u7528\u4e8eIgG3\u548cIgM\u578b\u5355\u6297\u7684\u63d0\u53d6\uff0c\u6240\u83b7\u5236\u54c1\u7684\u6297\u4f53\u6d3b\u6027\u51e0\u4e4e\u4fdd\u6301\u4e0d\u53d8\uff0c\u5bf9IgG3\u5355\u6297\u7684\u56de\u6536\u7387\u9ad8\u4e8e90%\uff0c\u5bf9IgM\u5355\u6297\u7684\u56de\u6536\u7387\u4e3a 40-90%\u4e0d\u7b49\u3002\u5176\u64cd\u4f5c\u6b65\u9aa4\u5982\u4e0b\uff1a\u53d6\u4e00\u5b9a\u91cf\u7684\u9884\u5904\u7406\u8fc7\u7684\u8179\u6c34\uff0c\u5148\u540e\u52a0\u5165NaCl\u548cCaCl2\uff0c\u4f7f\u5404\u81ea\u7684\u6d53\u5ea6\u5206\u522b\u8fbe0.2mol\/L\u548c25mmol\/L\uff0c\u968f\u4e4b\u53ef\u89c1\u7ea4\u7ef4\u86cb\u767d\u7684\u4ea7\u751f\uff1b\u7ecf\u6ee4\u7eb8\u8fc7\u6ee4\u540e\uff0c\u6ee4\u6db2\u5bf9100\u500d\u4f53\u79ef\u7684\u53bb\u79bb\u5b50\u6c34\u900f\u6790\uff0c4\u2103 8-15\u5c0f\u65f6\uff08\u82e5\u662fIgG3\u5355\u6297\uff0c\u4e5f\u53ef\u5ba4\u6e292\u5c0f\u65f6\uff09\uff0c\u5176\u95f4\u6362\u6c341-2\u6b21\uff1b\u53d6\u51fa\u540e22000g\u79bb\u5fc330\u5206\u949f\uff0c\u5f03\u4e0a\u6e05\uff1b\u5c06\u6c89\u6dc0\u6eb6\u4e8ePH8.0 1mol\/L NaCl\uff0c0.1mol\/L Tris-HCl\u6eb6\u6db2\u4e2d\uff0c\u91cd\u590d\u4e0a\u8ff0\u7684\u900f\u6790\u4e0e\u79bb\u5fc3\uff1b\u5c06\u6c89\u6dc0\u7684\u4f18\u7403\u86cb\u767d\u6d53\u5ea6\u8c03\u81f35-10mg\/ml\uff0c\u5206\u88c5\u51bb\u5b58\u5907\u7528\u3002<\/div>\n
\u4e09\u3001\u5355\u6297\u7eaf\u5316\u7684\u65b9\u6cd5<\/b><\/div>\n
\u5355\u6297\u7eaf\u5316\u7684\u65b9\u6cd5\u6709\u5f88\u591a\u79cd\uff0c\u5e94\u6839\u636e\u5177\u4f53\u5355\u6297\u7684\u7279\u6027\u548c\u5b9e\u9a8c\u6761\u4ef6\u9009\u62e9\u9002\u5b9c\u7684\u65b9\u6cd5\uff0c\u5e38\u7528\u7684\u6280\u672f\u6709DEAE\u79bb\u5b50\u4ea4\u6362\u5c42\u6790\u67f1\u3001\u51dd\u80f6\u8fc7\u6ee4\u6cd5\u548c\u4eb2\u548c\u5c42\u6790\u6cd5\u4e09\u79cd\u3002<\/div>\n","protected":false},"excerpt":{"rendered":"

\u4e00\u3001\u8179\u6c34\u578b\u5355\u6297\u7684\u7eaf\u5316 \u5728\u5355\u6297\u7eaf\u5316\u4e4b\u524d\uff0c\u4e00\u822c\u5747\u9700\u5bf9\u8179\u6c34\u8fdb\u884c\u9884\u5904\u7406\uff0c\u76ee\u7684\u662f\u4e3a\u4e86\u8fdb\u4e00\u6b65\u9664\u53bb\u7ec6\u80de\u53ca\u5176\u6b8b\u6e23\u3001\u5c0f\u9897\u7c92\u7269\u8d28\u3001 […]<\/p>\n","protected":false},"author":1,"featured_media":73,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":[],"categories":[5,4],"tags":[],"_links":{"self":[{"href":"http:\/\/chundubio.com\/index.php?rest_route=\/wp\/v2\/posts\/1738"}],"collection":[{"href":"http:\/\/chundubio.com\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"http:\/\/chundubio.com\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"http:\/\/chundubio.com\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"http:\/\/chundubio.com\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=1738"}],"version-history":[{"count":1,"href":"http:\/\/chundubio.com\/index.php?rest_route=\/wp\/v2\/posts\/1738\/revisions"}],"predecessor-version":[{"id":1739,"href":"http:\/\/chundubio.com\/index.php?rest_route=\/wp\/v2\/posts\/1738\/revisions\/1739"}],"wp:featuredmedia":[{"embeddable":true,"href":"http:\/\/chundubio.com\/index.php?rest_route=\/wp\/v2\/media\/73"}],"wp:attachment":[{"href":"http:\/\/chundubio.com\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=1738"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"http:\/\/chundubio.com\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=1738"},{"taxonomy":"post_tag","embeddable":true,"href":"http:\/\/chundubio.com\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=1738"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}