{"id":1598,"date":"2022-01-11T18:30:12","date_gmt":"2022-01-11T09:30:12","guid":{"rendered":"https:\/\/anatomy.med.gunma-u.ac.jp\/protocols\/?p=1598"},"modified":"2022-01-12T19:54:00","modified_gmt":"2022-01-12T10:54:00","slug":"%e5%85%8d%e7%96%ab%e6%9f%93%e8%89%b2","status":"publish","type":"post","link":"https:\/\/anatomy.med.gunma-u.ac.jp\/protocols\/?p=1598","title":{"rendered":"\u514d\u75ab\u67d3\u8272"},"content":{"rendered":"

\u9053\u5177
\n\u25cfFALCON 353047 24well\u3000\u7d30\u80de\u57f9\u990a\u7528\u30de\u30eb\u30c1\u30a6\u30a7\u30eb\u30d7\u30ec\u30fc\u30c8
\n\u203b\u7d30\u80de\u57f9\u990a\u7528\u3067\u3059\u304c\u5728\u5eab\u304c\u4f59\u3063\u3066\u3044\u308b\u306e\u3067\u4ee3\u7528\u3057\u3066\u3044\u307e\u3059
\n\u3082\u3057\u4f7f\u3044\u5207\u3063\u305f\u5834\u5408\u306f\u4ed6\u306e\u5ec9\u4fa1\u7248\u306e24well\u30d7\u30ec\u30fc\u30c8\u306b\u5207\u308a\u66ff\u3048\u307e\u3059
\n\u25cf\u30cd\u30a4\u30eb\u7528\u30d6\u30e9\u30b7 (\u30c0\u30a4\u30bd\u30fc\u30677\u672c\u3067100\u5186\u3067\u58f2\u3063\u3066\u3044\u308b)<\/p>\n

\u8a66\u85ac(\u4fdd\u5b58\u5834\u6240)
\nPBS (\u5ba4\u6e29\u307e\u305f\u306f4\u2103)
\n0.5% Triton-X100\/PBS (4\u2103)
\nNormal Donkey Serum (Jackson)<\/a> (-20\u2103)<\/p>\n

\u624b\u9806
\nDAY1<\/strong>
\n1. \u4f7f\u7528\u3059\u308bwell\u306b250\u03bcL\u306e0.5% Triton-X100\/PBS\u3092\u5165\u308c\u308b
\n2. \u30cd\u30a4\u30eb\u7528\u30d6\u30e9\u30b7\u3092\u4f7f\u3063\u3066\u5207\u7247\u3092\u63ac\u3063\u3066\u5165\u308c\u308b
\n3. \u5ba4\u6e29\u3067\u632f\u3068\u3046\u300130min (permeabilize)
\n4. NDS\u30925\u03bcL\u52a0\u3048\u308b(final 2%)
\n5. \u5ba4\u6e29\u3067\u632f\u3068\u3046\u300130min (blocking)
\n6. 1\u6b21\u6297\u4f53\u3092\u9069\u91cf\u52a0\u3048\u308b
\n7. \u5ba4\u6e29\u3067\u632f\u3068\u3046\u3001O\/N<\/p>\n

DAY2<\/strong>
\n1. PBS\u306b\u4ea4\u63db(\u6d17\u3073\u3093\u304b\u3089\u9069\u91cf)5min x 3
\n2. 2\u6b21\u6297\u4f53\u3092\u9069\u91cf\u52a0\u3048\u308b(2\u6b21\u6297\u4f53\u306fCan Get Signal\u306b\u5e0c\u91c8\u3059\u308b)
\n3. \u5ba4\u6e29\u3067\u632f\u3068\u3046\u300160min
\n4. PBS\u306b\u4ea4\u63db(\u6d17\u3073\u3093\u304b\u3089\u9069\u91cf)5min x 3
\n5. \u30b9\u30e9\u30a4\u30c9\u30ac\u30e9\u30b9\u306b\u5341\u5206\u91cf\u306ePBS\u3092\u5782\u3089\u3059
\n6. \u30cd\u30a4\u30eb\u7528\u30d6\u30e9\u30b7\u3092\u4f7f\u3063\u3066\u5207\u7247\u3092\u63ac\u3063\u3066\u30b9\u30e9\u30a4\u30c9\u30ac\u30e9\u30b9\u306b\u79fb\u3059
\n7. PBS\u3092\u5438\u3044\u306a\u304c\u3089\u5207\u7247\u306e\u4f4d\u7f6e\u3092\u8abf\u7bc0\u3059\u308b\u3002
\n8. \u5c01\u5165\u6db2\u30921\u6ef4\u5782\u3089\u3059
\n9. \u30ab\u30d0\u30fc\u30ac\u30e9\u30b9\u3092\u88ab\u305b\u308b
\n10. \u4f59\u5206\u306a\u5c01\u5165\u6db2\u3092\u30ad\u30e0\u30ef\u30a4\u30d7\u3067\u5438\u3046
\n\u3000\u3000\u2190\u3057\u3063\u304b\u308a\u3068\u5438\u308f\u306a\u3044\u3068\u3001\u30b9\u30e9\u30a4\u30c9G\u3068\u30ab\u30d0\u30fcG\u306b\u9699\u9593\u304c\u3067\u304d\u3066\u30d4\u30f3\u30c8\u304c\u5408\u308f\u306a\u3044\u3001\u53ce\u5dee\u306e\u539f\u56e0\u306b\u306a\u308b
\n11. \u30ab\u30d0\u30fc\u30ac\u30e9\u30b9\u306e\u7e01\u306b\u30cd\u30a4\u30eb\u30b3\u30fc\u30c8\u3092\u5857\u308b<\/p>\n","protected":false},"excerpt":{"rendered":"

\u9053\u5177 \u25cfFALCON 353047 24well\u3000\u7d30\u80de\u57f9\u990a\u7528\u30de\u30eb\u30c1\u30a6\u30a7\u30eb\u30d7\u30ec\u30fc\u30c8 \u203b\u7d30\u80de\u57f9\u990a\u7528\u3067\u3059\u304c\u5728\u5eab\u304c\u4f59\u3063\u3066\u3044\u308b\u306e\u3067\u4ee3\u7528\u3057\u3066\u3044\u307e\u3059 \u3082\u3057\u4f7f\u3044\u5207\u3063\u305f\u5834\u5408\u306f\u4ed6\u306e\u5ec9\u4fa1\u7248\u306e24well\u30d7\u30ec\u30fc\u30c8\u306b\u5207\u308a\u66ff\u3048\u307e\u3059 \u25cf\u30cd\u30a4\u30eb\u7528\u30d6\u30e9\u30b7<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"inline_featured_image":false,"_jetpack_newsletter_access":"","_jetpack_dont_email_post_to_subs":false,"_jetpack_newsletter_tier_id":0,"_jetpack_memberships_contains_paywalled_content":false,"_jetpack_memberships_contains_paid_content":false,"footnotes":"","jetpack_publicize_message":"","jetpack_publicize_feature_enabled":true,"jetpack_social_post_already_shared":true,"jetpack_social_options":{"image_generator_settings":{"template":"highway","default_image_id":0,"font":"","enabled":false},"version":2},"jetpack_post_was_ever_published":false},"categories":[10],"tags":[],"class_list":["post-1598","post","type-post","status-publish","format-standard","hentry","category-cell-biology"],"jetpack_publicize_connections":[],"jetpack_featured_media_url":"","jetpack_sharing_enabled":true,"jetpack_shortlink":"https:\/\/wp.me\/p4fFyn-pM","jetpack-related-posts":[],"_links":{"self":[{"href":"https:\/\/anatomy.med.gunma-u.ac.jp\/protocols\/index.php?rest_route=\/wp\/v2\/posts\/1598","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/anatomy.med.gunma-u.ac.jp\/protocols\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/anatomy.med.gunma-u.ac.jp\/protocols\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/anatomy.med.gunma-u.ac.jp\/protocols\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/anatomy.med.gunma-u.ac.jp\/protocols\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=1598"}],"version-history":[{"count":6,"href":"https:\/\/anatomy.med.gunma-u.ac.jp\/protocols\/index.php?rest_route=\/wp\/v2\/posts\/1598\/revisions"}],"predecessor-version":[{"id":1607,"href":"https:\/\/anatomy.med.gunma-u.ac.jp\/protocols\/index.php?rest_route=\/wp\/v2\/posts\/1598\/revisions\/1607"}],"wp:attachment":[{"href":"https:\/\/anatomy.med.gunma-u.ac.jp\/protocols\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=1598"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/anatomy.med.gunma-u.ac.jp\/protocols\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=1598"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/anatomy.med.gunma-u.ac.jp\/protocols\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=1598"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}