{"id":4078,"date":"2023-07-11T10:40:54","date_gmt":"2023-07-11T02:40:54","guid":{"rendered":"https:\/\/www.haokebio.com\/?p=4078"},"modified":"2024-10-10T14:24:28","modified_gmt":"2024-10-10T06:24:28","slug":"%e9%ac%bc%e7%ac%94%e7%8e%af%e8%82%bd%ef%bc%88%e7%bb%bf%e5%85%89%ef%bc%89","status":"publish","type":"post","link":"https:\/\/www.haokebio.com\/4078.html","title":{"rendered":"\u9b3c\u7b14\u73af\u80bd\uff08\u7eff\u5149\uff09"},"content":{"rendered":"\n<section class=\"wp-block-themepark-block-themepark-area block_layout thempark-block undefined\" id=\"\" style=\"padding-bottom:14px;padding-top:28px;\" data-swiper-parallax=\"-700\"><div class=\"block_layout_in\" style=\"\">\n<div class=\"wp-block-column block_layout_in\">\n<div class=\"wp-block-themepark-block-themepark-product-head content_pro bing undefined\" style=\"background-color:#ffffff;margin-bottom:20px;\"><div class=\"tab_pic\" style=\"width:47%\"><div class=\"left_post_pic\"><a class=\" swipebox\" href=\"https:\/\/www.haokebio.com\/wp-content\/uploads\/2023\/07\/hki0018.png?imageView2\/0\/format\/webp\/q\/75\"><img decoding=\"async\" class=\"lazy lazyload\" data-src=\"https:\/\/www.haokebio.com\/wp-content\/uploads\/2023\/07\/hki0018.png?imageView2\/0\/format\/webp\/q\/75\" src=\"https:\/\/www.haokebio.com\/wp-content\/themes\/smartenterprise\/images\/loading.png\" alt=\"\u9b3c\u7b14\u73af\u80bd\uff08\u7eff\u5149\uff09\"\/><\/a><\/div><\/div><div class=\"ds\" style=\"width:48%\">\n<div class=\"wp-block-column\"><header class=\"post_title_head \" style=\"margin-bottom:7px;\"><div class=\"title\"><h1 class=\"h1title\" style=\"font-size:24px;\">\u9b3c\u7b14\u73af\u80bd\uff08\u7eff\u5149\uff09<\/h1><\/div><div class=\"description\"><span ><i class=\"fa fa-folder\"><\/i><a  href=\"https:\/\/www.haokebio.com\/category\/products\/cpzh\/zhother\/\">\u5176\u4ed6<\/a> <a  href=\"https:\/\/www.haokebio.com\/category\/products\/cpzh\/\">\u5206\u5b50\u75c5\u7406<\/a> <a  href=\"https:\/\/www.haokebio.com\/category\/products\/\">\u8bd5\u5242<\/a> <\/span><\/div><\/header><ul class=\"csbox_pt \"  ><li class=\"jgss\"style=\"border-bottom:1px solid #e0e0e0;\"><span style=\"font-size:14px;color:#666666;width:125px;\">\u4ef7\u683c<\/span><span style=\"font-size:16px;color:#e21f2f;font-weight: bold;\">\u00a52050<\/span><\/li><li style=\"border-bottom:1px solid #e0e0e0;\"><span style=\"font-size:14px;color:#666666;width:125px;\">\u89c4\u683c<\/span><span style=\"color:#666666;\">300T<\/span><\/li><li style=\"border-bottom:1px solid #e0e0e0;\"><span style=\"font-size:14px;color:#666666;width:125px;\">\u8d27\u53f7<\/span><span style=\"color:#666666;\">HKI0018G<\/span><\/li><li style=\"border-bottom:1px solid #e0e0e0;\"><span style=\"font-size:14px;color:#666666;width:125px;\">\u5355\u4f4d<\/span><span style=\"color:#666666;\">\u74f6<\/span><\/li><li style=\"border-bottom:1px solid #e0e0e0;\"><span style=\"font-size:14px;color:#666666;width:125px;\">\u54c1\u724c<\/span><span style=\"color:#666666;\">HaoKebio<\/span><\/li><\/ul>\n\n\n<div class=\"wp-block-buttons\">\n<div class=\"wp-block-button has-custom-width wp-block-button__width-75 has-custom-font-size has-normal-font-size\"><a class=\"wp-block-button__link has-monte-white-color has-luminous-vivid-orange-background-color has-text-color has-background wp-element-button\" style=\"border-radius:10px\" rel=\"http:\/\/wpa.qq.com\/msgrd?v=3&amp;uin=53383090&amp;site=qq&amp;menu=yes\">QQ53383090 \u54a8\u8be2\u8d2d\u4e70<\/a><\/div>\n<\/div>\n\n\n\n<p><\/p>\n<\/div>\n<\/div><\/div>\n<\/div>\n<\/div><div class=\"layout_badh\"><div class=\"shizhezhao\" style=\"opacity:0.85;background-color:#ffffff;\"><\/div><div class=\"PcOnly themepark-block_layout_ba\"><video id=\"VideoPlayer\" src=\"http:\/\/data.themepark.com.cn\/2021\/n\/smartenterprise_data\/qy13\/uploads\/2022\/05\/ev_20220524_140257.mp4\" autoplay loop muted defaultmuted=\"\" playsinline><source src=\"http:\/\/data.themepark.com.cn\/2021\/n\/smartenterprise_data\/qy13\/uploads\/2022\/05\/ev_20220524_140257.mp4\" type=\"video\/mp4\"\/><\/video><\/div><\/div><\/section>\n\n\n\n<div class=\"wp-block-themepark-block-themepark-tab thempark-block thempark_tab_box undefined\" style=\"margin-bottom:20px;\" bloop=\"no\" auto=\"no\" mouse=\"click\"><div class=\"content_tab_title\" style=\"background-color:#eaeaea;\"><div class=\"fidxtab\" style=\"background-color:#eaeaea;\"><div class=\"btn active  tabcolor5  \" style=\"\"><span class=\"tabtt\" style=\"\"><i class=\"fas fa fa-fas fa-clipboard-check\" style=\"\"><\/i>\u4ea7\u54c1\u4ecb\u7ecd<\/span><\/div><div class=\"btn tabcolor5  \" style=\"\"><span class=\"tabtt\" style=\"\"><i class=\"fas fa fa-fas fa-charging-station\" style=\"\"><\/i>\u4ea7\u54c1\u53c2\u6570<\/span><\/div><div class=\"btn tabcolor5  \" style=\"\"><span class=\"tabtt\" style=\"\"><i class=\"fas fa fa-fas fa-file-pdf\" style=\"\"><\/i>\u8d44\u6599\u4e0b\u8f7d<\/span><\/div><\/div><\/div><div class=\"swiper-container content_tab_swiper\" style=\"background-color:#ffffff;\"><div class=\"swiper-wrapper\">\n<div class=\"wp-block-column ecdorbox0 swiper-slide tap_s-box active\" style=\"font-size:15px\">\n<figure class=\"wp-block-table\"><table><tbody><tr><th>\u54c1\u724c<\/th><th>\u5546\u54c1\u7f16\u7801<\/th><th>\u5546\u54c1\u540d\u79f0<\/th><th>\u89c4\u683c<\/th><th>\u5355\u4f4d<\/th><th>\u4ef7\u683c<\/th><\/tr><tr><th><span style=\"color: #333333;\">HaoKebio<\/span><\/th><th>HKI0018G<\/th><th>\u9b3c\u7b14\u73af\u80bd\uff08\u7eff\u5149\uff09<\/th><th>300ul<\/th><th>\u74f6<\/th><th>\uffe52050<\/th><\/tr><\/tbody><\/table><\/figure>\n\n\n\n<p><strong>\u3010\u4ea7\u54c1\u7b80\u4ecb\u3011<\/strong><\/p>\n\n\n\n<p>\u9b3c\u7b14\u73af\u80bd\uff08Phalloidin\uff09\uff0c\u53c8\u79f0\u9b3c\u7b14\u9e45\u818f\u7d20\uff0c\u6700\u521d\u662f\u4ece\u6bd2\u8611\u83c7\u9b3c\u7b14\u9e45\u818f\u83cc(Amanitaphalloides)\u4e2d\u5206\u79bb\u5230\u7684\u4e00\u79cd\u53cc\u73af\u80bd\uff0c\u53ef\u4ee5\u6781\u9ad8\u7684\u4eb2\u548c\u529b\u548c\u7279\u5f02\u6027\u4e0e\u808c\u52a8\u86cb\u767d\u4e1d F-actin \u7ed3\u5408\uff0c\u4e0d\u4f1a\u7ed3\u5408\u5355\u4f53\u808c\u52a8\u86cb\u767d\uff08G-actin\uff09\u3002\u9b3c\u7b14\u73af\u80bd\u5bf9\u5927\u5c0f\u7ea4\u7ef4\u7684\u4eb2\u548c\u529b\u76f8\u8fd1\uff0c\u5728\u8bb8\u591a\u4e0d\u540c\u7684\u52a8\u690d\u7269\u7269\u79cd\u7684\u808c\u8089\u548c\u975e\u808c\u8089\u7ec6\u80de\u4e2d\uff0c\u57fa\u672c\u90fd\u6309\u7167\u4e00\u4e2a\u808c\u52a8\u86cb\u767d\u4e9a\u57fa\u4e0e\u4e00\u4e2a\u9b3c\u7b14\u73af\u80bd\u5206\u5b50\u7684\u5316\u5b66\u8ba1\u91cf\u6bd4\u7ed3\u5408\u3002\u4e0d\u50cf\u808c\u52a8\u86cb\u767d\u6297\u4f53\uff0c\u5bf9\u4e0d\u540c\u7269\u79cd\u6216\u6765\u6e90\u7684\u673a\u52a8\u86cb\u767d\u4eb2\u548c\u529b\u4f1a\u53d1\u751f\u660e\u663e\u53d8\u5316\u3002\u9b3c\u7b14\u73af\u80bd\u7684\u975e\u7279\u5f02\u6027\u7ed3\u5408\u51e0\u4e4e\u53ef\u4ee5\u5ffd\u7565\uff0c\u67d3\u8272\u548c\u672a\u67d3\u8272\u533a\u57df\u7684\u5dee\u5f02\u975e\u5e38\u660e\u663e\u3002\u9b3c\u7b14\u73af\u80bd\u53ca\u5176\u884d\u751f\u7269\u5728\u7eb3\u6469\u5c14\u6d53\u5ea6\u5373\u53ef\u5bf9 F-actin \u67d3\u8272\uff0c\u53ef\u4ee5\u975e\u5e38\u65b9\u4fbf\u7684\u6807\u8bb0\u3001\u8bc6\u522b\u548c\u5b9a\u91cf\u7814\u7a76 F-actin \u7684\u5206\u5e03\u3002\u672c\u4ea7\u54c1\u4e3a\u8367\u5149\u67d3\u6599 FITC \u6807\u8bb0\u7684\u9b3c\u7b14\u73af\u80bd\uff0c\u4ee5 20 \u03bcM \u50a8\u5b58\u6db2\u5f62\u5f0f\u63d0\u4f9b\uff0c\u6eb6\u5242\u4e3a\u7532\u9187\u3002\u5e38\u7528\u6d53\u5ea6\u8303\u56f4 80-200nM\uff0c\u6bcf\u6b21\u67d3\u8272\u4f7f\u7528 200 \u03bcL \u5de5\u4f5c\u6db2\u5373\u53ef\u3002<\/p>\n\n\n\n<p><strong>\u3010\u50a8\u5b58\u4e0e\u8fd0\u8f93\u3011<\/strong><\/p>\n\n\n\n<p>\u51b0\u888b\uff08wet ice\uff09\u8fd0\u8f93\uff0c-20\u2103\u907f\u5149\u4fdd\u5b58\uff1b\u6709\u6548\u671f 12 \u4e2a\u6708\u3002<\/p>\n\n\n\n<p><strong>\u3010\u4f7f\u7528\u65b9\u6cd5\u3011<\/strong><\/p>\n\n\n\n<p><strong>\u8bd5\u5242\u51c6\u5907\uff1a<\/strong>\u81ea\u5907 1\u00d7PBS \u7f13\u51b2\u6db2\uff08pH 7.2-7.4\uff09\uff0c\u56fa\u5b9a\u6db2\uff08\u8d27\u53f7\uff1aHK2003\uff09\uff0c\u7ec6\u80de\u901a\u900f\u5242\uff08\u8d27\u53f7\uff1aHKI0048\uff09\uff0c\u6297\u8367\u5149\u6dec\u706d\u5c01\u7247\u5242\uff08\u8d27\u53f7\uff1aHKI0007\uff09\uff0c\u5373\u7528\u578b DAPI \u67d3\u8272\u6db2\uff08\u8d27\u53f7\uff1aHKI0005\uff09\u7b49\u3002<\/p>\n\n\n\n<p><strong>\u5de5\u4f5c\u6db2\u914d\u5236\uff1a<\/strong>\u7528 1\u00d7PBS \u5c06\u672c\u4ea7\u54c1\u7a00\u91ca\u5230\u9700\u8981\u7684\u5de5\u4f5c\u6d53\u5ea6\uff0c\u5e38\u7528\u5de5\u4f5c\u6d53\u5ea6\u4e3a 80-200 nM\u3002\u63a8\u8350\u4f7f\u7528 100nM\uff0c\u5373\u6bcf 1 \u03bcL FITC \u6807\u8bb0\u9b3c\u7b14\u73af\u80bd\u4e0e 199 \u03bcL PBS \u6df7\u5300\u3002\u6700\u4f73\u67d3\u8272\u5de5\u4f5c\u6d53\u5ea6\u8bf7\u53c2\u8003\u6587\u732e\u6216\u8fdb\u884c\u9884\u5b9e\u9a8c\u6478\u7d22\u3002\u9b3c\u7b14\u73af\u80bd\u67d3\u8272\u5de5\u4f5c\u6db2\uff0c\u73b0\u914d\u73b0\u7528\uff0c\u5ba4\u6e29\u907f\u5149\u4fdd\u5b58\u3002<\/p>\n\n\n\n<p><strong>\u64cd\u4f5c\u6b65\u9aa4<\/strong><\/p>\n\n\n\n<p>1. \u57f9\u517b\u597d\u7684\u7ec6\u80de\u722c\u7247\uff08\u5bc6\u5ea6\u81f3\u5c11\u8fbe\u5230\u534a\u6c47\u5408\uff09\uff0c\u53bb\u9664\u57f9\u517b\u6db2\uff0c\u7528 37\u2103\u9884\u70ed\u7684 1\u00d7PBS \u7f13\u51b2\u6db2\u6e05\u6d17\u7ec6\u80de\u6e05\u6d17\u7ec6\u80de 2 \u6b21\u3002<\/p>\n\n\n\n<p>2. \u52a0\u5165\u542b 4%\u7532\u919b\u7684\u56fa\u5b9a\u6db2\u8986\u76d6\u7ec6\u80de\uff0c\u5ba4\u6e29\u56fa\u5b9a 10 min\u3002\u6ce8\u610f\uff0c\u7532\u9187\u80fd\u7834\u574f\u808c\u52a8\u86cb\u767d\uff0c\u56e0\u6b64\u56fa\u5b9a\u6db2\u4e2d\u4e0d\u5f97\u542b\u6709\u7532\u9187\uff0c\u907f\u514d\u4f7f\u7528\u542b\u7532\u9187\u7684\u7532\u919b\u6eb6\u6db2\u3002<\/p>\n\n\n\n<p>3. \u7528 1\u00d7PBS \u7f13\u51b2\u6db2\u6e05\u6d17\u7ec6\u80de 2-3 \u6b21\uff0c\u6bcf\u6b21 30 s-1 min\u3002<\/p>\n\n\n\n<p>4. \u7528\u7ec6\u80de\u901a\u900f\u5242\u8986\u76d6\u7ec6\u80de\uff0c\u5ba4\u6e29\u900f\u5316\u7ec6\u80de 5-10 min\u3002<\/p>\n\n\n\n<p>5. \u7528 1\u00d7PBS \u7f13\u51b2\u6db2\u6e05\u6d17\u7ec6\u80de 2-3 \u6b21\uff0c\u6bcf\u6b21 30 s-1 min\u3002<\/p>\n\n\n\n<p>6. \u7ec6\u80de\u722c\u7247\u7a0d\u5fae\u7529\u5e72\u540e\uff0c\u7528\u7ec4\u5316\u7b14\u753b\u5708\u4f7f\u7ec6\u80de\u4f4d\u4e8e\u5708\u4e2d\u592e\u3002\u53d6 200 \u03bcL \u73b0\u914d\u7684 FITC \u6807\u8bb0\u9b3c\u7b14\u73af\u80bd\u67d3\u8272\u5de5\u4f5c\u6db2\u5b8c\u5168\u8986\u76d6\u7ec6\u80de\uff0c\u5ba4\u6e29\u907f\u5149\u5b75\u80b2 30 min\u3002\u6ce8\u610f\uff0c\u901a\u5e38\u60c5\u51b5\u4e0b\uff0c4-37\u2103\u5747\u9002\u5408\u67d3\u8272\u3002\u4e3a\u907f\u514d\u67d3\u8272\u5de5\u4f5c\u6db2\u6325\u53d1\u5bfc\u81f4\u5e72\u7247\uff0c\u5b75\u80b2\u8fc7\u7a0b\u9700\u5c06\u7ec6\u80de\u722c\u7247\u7f6e\u4e8e\u5bc6\u5c01\u5bb9\u5668\u5185\uff0c\u4f8b\u5982\u907f\u5149\u6e7f\u76d2\u3002<\/p>\n\n\n\n<p>7. \u7528 1\u00d7PBS \u7f13\u51b2\u6db2\u6e05\u6d17\u7ec6\u80de 2-3 \u6b21\uff0c\u6bcf\u6b21 30 s-1 min\u3002<\/p>\n\n\n\n<p>8. \uff08\u53ef\u9009\u505a\uff09\u5411\u722c\u7247\u4e0a\u6ef4\u52a0\u5373\u7528\u578b DAPI \u67d3\u8272\u6db2\u8986\u76d6\u7ec6\u80de\uff0c\u8fdb\u884c\u7ec6\u80de\u6838\u67d3\u8272 3-5 min\u3002\u75281\u00d7PBS \u7f13\u51b2\u6db2\u6e05\u6d17\u7ec6\u80de 2-3 \u6b21\uff0c\u6bcf\u6b21 30 s-1 min<\/p>\n\n\n\n<p>9. \u7ec6\u80de\u722c\u7247\u7a0d\u7529\u5e72\u540e\uff0c\u5012\u7f6e\u5728\u6ef4\u52a0\u4e00\u6ef4\u6297\u8367\u5149\u6dec\u706d\u5c01\u7247\u5242\u7684\u8f7d\u73bb\u7247\u4e0a\uff0c\u7528\u7eb8\u5dfe\u5438\u6389\u5bf9\u4e8e\u5c01\u7247\u5242\u3002<\/p>\n\n\n\n<p>\u3010\u53ef\u9009\u6b65\u9aa4\u3011\uff1a\u5b8c\u6210\u6b65\u9aa4 7 \u540e\uff0c\u76f4\u63a5\u5c06\u76d6\u73bb\u7247\u5012\u7f6e\u5728\u6ef4\u52a0\u542b DAPI \u7684\u6297\u8367\u5149\u6dec\u706d\u5c01\u7247\u5242\uff08\u8d27\u53f7\uff1aHKI0007-2\uff09\u7684\u8f7d\u73bb\u7247\u4e0a\uff0c\u7136\u540e\u5438\u6389\u591a\u4f59\u7684\u5c01\u7247\u5242\u3002\u4ee5\u8367\u5149\u663e\u5fae\u955c\u6216\u6fc0\u5149\u5171\u805a\u7126\u663e\u5fae\u955c\u89c2\u5bdf\u7ed3\u679c\u3002<\/p>\n\n\n\n<p>10. \u8367\u5149\u663e\u5fae\u955c\u6216\u6fc0\u5149\u5171\u805a\u7126\u663e\u5fae\u955c\u89c2\u5bdf\u7ed3\u679c\uff0c\u9009\u62e9 FITC\uff08Ex\/Em=492\/518 nm\uff09\u548c DAPI\uff08Ex\/Em=364\/454nm\uff09\u901a\u9053\u3002<\/p>\n\n\n\n<p><strong>\u3010\u6ce8\u610f\u4e8b\u9879\u3011<\/strong><\/p>\n\n\n\n<p>1. \u8367\u5149\u67d3\u6599\u90fd\u5b58\u5728\u6dec\u706d\u7684\u95ee\u9898\uff0c\u5efa\u8bae\u67d3\u8272\u540e\u5c3d\u5feb\u5b8c\u6210\u68c0\u6d4b\u53ca\u89c2\u5bdf\u62cd\u7167\u3002<\/p>\n\n\n\n<p>2. \u7532\u9187\u53ef\u4ee5\u7834\u574f actin \u86cb\u767d\uff0c\u56e0\u6b64\u6837\u672c\u524d\u671f\u56fa\u5b9a\u4e0d\u80fd\u4f7f\u7528\u542b\u6709\u7532\u9187\u7684\u56fa\u5b9a\u6db2\u3002<\/p>\n\n\n\n<p>3. \u9b3c\u7b14\u73af\u80bd\u901a\u5e38\u4e0d\u5177\u6709\u7ec6\u80de\u901a\u900f\u6027\uff0c\u6781\u5c11\u7528\u4e8e\u6d3b\u7ec6\u80de\u67d3\u8272\u3002<\/p>\n\n\n\n<p>4. \u672c\u4ea7\u54c1\u6eb6\u5242\u4e3a\u7532\u9187\uff0c\u6781\u6613\u6325\u53d1\uff0c\u6bcf\u6b21\u7528\u5b8c\u4ee5\u540e\u9700\u53ca\u65f6\u62e7\u7d27\u7ba1\u76d6\u5bc6\u5c01\u4fdd\u5b58\u3002<\/p>\n\n\n\n<p>5. 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