{"id":42663,"date":"2023-05-25T12:53:26","date_gmt":"2023-05-25T04:53:26","guid":{"rendered":"https:\/\/biopioneer.com.tw\/?p=42663"},"modified":"2023-07-11T15:38:59","modified_gmt":"2023-07-11T07:38:59","slug":"%e5%85%a7%e7%9a%ae-transwell-%e9%80%9a%e9%80%8f%e6%80%a7%e6%aa%a2%e6%b8%ac%e8%a9%a6%e5%8a%91%e7%9b%92-endothelial-transwell-permeability-assay-kit%e8%b2%a8%e8%99%9fcb6929","status":"publish","type":"post","link":"https:\/\/biopioneer.com.tw\/?p=42663","title":{"rendered":"\u5167\u76ae Transwell \u901a\u900f\u6027\u6aa2\u6e2c\u8a66\u5291\u76d2| Endothelial Transwell Permeability Assay Kit\u8ca8\u865fCB6929"},"content":{"rendered":"

\u5167\u76ae Transwell \u901a\u900f\u6027\u6aa2\u6e2c\u8a66\u5291\u76d2-Endothelial Transwell Permeability Assay Kit\u8ca8\u865fCB6929<\/span><\/strong><\/a><\/span><\/p>\n

Endothelial Transwell Permeability Assay Kit (24 SPL PC inserts, 100 assays) Catalog Number: CB6929<\/p>\n

Application<\/strong><\/span><\/p>\n

\u5b9a\u4f4d\u65bc\u8840\u7ba1\u58c1\u95dc\u9375\u754c\u9762\u7684\u5fae\u8840\u7ba1\u5167\u76ae\u7d30\u80de\u55ae\u5c64(microvascular endothelial cell monolayer )\u5177\u6709\u8abf\u7bc0\u7d44\u7e54\u6db2\u5e73\u8861( tissue fluid balanc)\u548c\u63d0\u4f9b\u6a5f\u9ad4\u751f\u5b58\u6240\u9700\u7684\u5fc5\u9700\u71df\u990a\u7d20(essential nutrients)\u7684\u91cd\u8981\u529f\u80fd\u3002\u5167\u76ae\u7d30\u80de(endothelial cells )\u80fd\u5920\u52d5\u614b\u8abf\u7bc0\u5176\u7528\u65bc\u904b\u8f38\u8840\u6f3f\u86cb\u767d(transport of plasma proteins)\u3001\u6eb6\u8cea(solutes)\u548c\u6db2\u9ad4\u7684\u7d30\u80de\u65c1\u8def\u548c\u8de8\u7d30\u80de\u901a\u8def\u3002\u5167\u76ae\u5c4f\u969c( endothelial barrier)\u662f\u4e00\u500b\u8abf\u7bc0\u826f\u597d\u7684\u7d50\u69cb\uff0c\u5b83\u5728\u6b63\u5e38\u751f\u7406\u689d\u4ef6\u4e0b\u5c0d\u6db2\u9ad4\u548c\u5206\u5b50\u4fdd\u6301\u6700\u5c0f\u548c\u9078\u64c7\u6027\u7684\u6ef2\u900f\u6027\u3002\u8840\u7ba1\u6ef2\u6f0f\u662f\u591a\u7a2e\u75be\u75c5\u7684\u91cd\u8981\u7279\u5fb5\uff0c\u4f8b\u5982\u611f\u67d3\u6027\u4f11\u514b(septic shock)\u3001\u708e\u75c7(inflammation)\u3001\u8840\u8166\u5c4f\u969c (blood-brain barrier,BBB) \u76f8\u95dc\u795e\u7d93\u7cfb\u7d71\u75be\u75c5\u7684\u7834\u58de\u3001\u75c5\u6bd2\u6027\u51fa\u8840\u71b1(viral hemorrhagic fever)\u3001\u764c\u75c7\u548c\u7f3a\u8840\u518d\u704c\u6ce8\u640d\u50b7\u3002\u8feb\u5207\u9700\u8981\u958b\u767c\u4e00\u7a2e\u9ad8\u5ea6\u9748\u654f\u548c\u53ef\u9760\u7684\u6aa2\u6e2c\u65b9\u6cd5\u4f86\u6e2c\u91cf\u5167\u76ae\u901a\u900f\u6027(endothelial permeability)\u3002 Cell Biologics<\/strong><\/span>\u958b\u767c\u4e86\u4e00\u7a2e\u7c21\u55ae\u3001\u53ef\u91cd\u8907\u3001\u9748\u654f\u3001\u5b9a\u91cf\u7684 transwell \u901a\u900f\u6027\u6e2c\u5b9a\u6cd5\u4f86\u6e2c\u91cf\u5167\u76ae\u5c4f\u969c\u529f\u80fd(endothelial barrier function )\uff0c\u4ee5\u4fbf\u5728\u9ad4\u5916\u5c0d\u591a\u7a2e\u985e\u578b\u7684\u5167\u76ae\u901a\u900f\u6027( endothelial permeability )\u9032\u884c\u5ee3\u6cdb\u5206\u6790\u3002\u8a72\u6e2c\u5b9a\u5c07\u5728\u5167\u76ae\u8655\u7406\u5f8c\u4e00\u5c0f\u6642\u5167\u901a\u904e\u57fa\u65bc\u5149\u8b5c\u5100\u7684\u5438\u5149\u5ea6\u8b80\u6578\u5668\uff08ELISA \u677f\u8b80\u6578\u5668\uff09\u5728 450 nm \u8655\u9032\u884c\u3002\u8a72\u6aa2\u6e2c\u8a66\u5291\u76d2\u9069\u7528\u65bc\u4f86\u81ea\u591a\u500b\u7269\u7a2e\u7684\u521d\u4ee3\u5167\u76ae\u7d30\u80de\u548c\u6c38\u751f\u7d30\u80de\u682a\u3002<\/p>\n

<\/p>\n

Storage Condition<\/strong><\/span><\/p>\n

Storage Temperature: 4 \u00b0C. Kit Components (for 100 assays in 96 well plates): A: SPL PC membrane, 6.5 mm, 0.4 \u00b5m pore, white, sterile 24 inserts in a 24-well cell culture plate. B: A 96-well ELISA plate (clear polystyrene wells, flat bottom). C: Streptavidin-HRP solution (150 \u00b5l). D: TMB substrate solution (5 ml). E: Stop solution (5 ml)<\/p>\n

Procedure All samples and standards should be run in triplicate.<\/p>\n

1. Add 1 ml endothelial growth medium in the experiment wells of 24-well plate.<\/p>\n

2. About 1~5 x 105 endothelial cells with or without transfection were seeded on the 6.5 mm transwell insert membrane (0.4 \u00b5l) in 200~400 \u00b5l endothelial growth medium.<\/p>\n

3. Transfer the inserts to the wells containing 1 ml of growth medium using a sterilized forceps and cover the plate.<\/p>\n

4. Incubate the plate in a cell culture incubator for 2 to 4 days until cells grow by 100% confluence. To check the cell confluence, move the inserts to the empty wells with a sterilized forceps, and wait for 5 minutes to check whether the medium from the upper inserts leaks into the wells of 24-well plate. If there is no leak, then proceed to the next steps. Otherwise, the cells need to grow for days till the cells grow a confluent monolayer.<\/p>\n

5. Endothelial cells were starved in a serum-free or 0.1% serum medium for a proper time according to individual experiment purpose.<\/p>\n

6. Add the desired stimulus (TNF-\uf061, IL-1\u03b2, thrombin, LPS, etc.) and 5 \u00b5l of streptavidin-HRP to the 300 \u00b5l fresh serum-free medium in the top insert chambers.<\/p>\n

7. Add 1 ml serum free medium in the wells of a new 24-well cell culture plate and transfer the inserts in to incubate in the 37 oC incubator for a proper treatment time. This assay works best when the cells are treated between 30 min to 24 hours.<\/p>\n

8. Transfer 20 \u00b5l of media from the lower chamber to the wells of 96-well ELISA assay plate. Each sample would be at least aliquoted in triplicate. No treatment or HRP-free medium were considered as negative control.<\/p>\n

9. Add 50 \u00b5l TMB substrate into each well of 96-well plate, incubate in a shaker for a couple of minutes at room temperature until the blue color occurred.<\/p>\n

10. Add 50 \u00b5l stop solution into the wells of 96-well plate, and the blue color will become yellow. Measure the spectrum absorption at 450 nm using an ELISA plate reader. Data expressed as A450 absorption readings are considered a relative permeability.<\/p>\n

<\/p>\n

\"\u7db2\u7ad9\u898f\u683c\u5c0f\u5716_\u5de5\u4f5c\u5340\u57df<\/a><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
Cat. #<\/td>\nName<\/td>\n<\/tr>\n
R3382<\/a><\/td>\n1X Red Blood Cell lysis buffer<\/a><\/td>\n<\/tr>\n
6920<\/a><\/td>\nAntibiotic\/Antimycotic Solution (1:100)<\/a><\/td>\n<\/tr>\n
6916<\/a><\/td>\nCell Culture Freezing Medium – ready to use<\/a><\/td>\n<\/tr>\n
6953<\/a><\/td>\nCollagen Type I, Rat Tail Solution – ready to use<\/a><\/td>\n<\/tr>\n
1166<\/a><\/td>\nEndothelial Cell Growth Supplement- ECGS (500X)<\/a><\/td>\n<\/tr>\n
6912<\/a><\/td>\nFetal Bovine Serum<\/a><\/td>\n<\/tr>\n
6950<\/a><\/td>\nGelatin-Based Coating Solution- ready to use<\/a><\/td>\n<\/tr>\n
3368<\/a><\/td>\nMacrophage Culture Supplement\/ L-cell supernatant<\/a><\/td>\n<\/tr>\n
6917<\/a><\/td>\nPenicillin\/Streptomycin Solution (1:100)<\/a><\/td>\n<\/tr>\n
6951<\/a><\/td>\nPoly-L-Lysine Solution<\/a><\/td>\n<\/tr>\n
6915<\/a><\/td>\nTrypsin\/EDTA 0.05% Solution<\/a><\/td>\n<\/tr>\n
6914<\/a><\/td>\nTrypsin\/EDTA 0.25% Solution<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

\"Cell<\/p>\n

Authorized Uses of\u00a0Cell Biologics<\/em>\u00a0Products<\/strong><\/span><\/p>\n

Endothelial Cell Growth Supplement from\u00a0Cell Biologics<\/em>\u00a0are distributed for\u00a0internal\u00a0in vitro<\/em>\u00a0research purposes only. Our products are not authorized for human use, for\u00a0in vitro<\/em>\u00a0diagnostic procedures, or for therapeutic procedures. Transfer or resale of any\u00a0Cell Biologics\u2019<\/em>\u00a0cells or products from the purchaser to other markets, organizations or individuals is prohibited by\u00a0Cell Biologics,<\/em>\u00a0without the company\u2019s written consent.\u00a0Cell Biologics\u2019<\/em>\u00a0Terms and Conditions must be accepted before submitting an order.<\/p>\n

\"\"<\/p>\n

Cell Biologics \u662f\u521d\u4ee3\u57f9\u990a\u7d30\u80de(\u00a0primary cultured cells)\u548c\u7d30\u80de\u57f9\u990a\u7522\u54c1\u7684\u4e3b\u8981\u88fd\u9020\u5546(cell culture products)\u3002\u63d0\u4f9b\u7bc4\u570d\u5ee3\u6cdb\u7684\u9ad8\u8cea\u91cf\u4eba\u985e\u548c\u52d5\u7269\u521d\u4ee3\u7d30\u80de\uff0c\u5305\u62ec\u5167\u76ae\u7d30\u80de(endothelial)\u3001\u4e0a\u76ae\u7d30\u80de(\u00a0epithelial)\u3001\u816b\u7624\u7d30\u80de(\u00a0tumor )\u548c\u5e79\u7d30\u80de(stem cells)\uff0c\u4ee5\u53ca\u512a\u5316\u7684\u7d30\u80de\u57f9\u990a\u57fa(optimized cell culture media)\u548c\u5176\u4ed6\u76f8\u95dc\u7522\u54c1\u3002\u5c0f\u9f20\u521d\u4ee3\u5167\u76ae\u7d30\u80de\u6700\u521d\u7531 Cell Biologics \u63d0\u4f9b\uff0c\u53ef\u6839\u64da\u7279\u5b9a\u79d1\u5b78\u9700\u6c42\u5b9a\u5236\u3002\u60a8\u53ef\u4ee5\u4fe1\u8cf4\u6211\u5011\u9ad8\u8cea\u91cf\u7684\u5b9a\u5236\u7d30\u80de\u5206\u96e2\u670d\u52d9\uff01\u8a72\u516c\u53f8\u9084\u5728\u958b\u767c\u91dd\u5c0d\u708e\u75c7\u6027\u75be\u75c5\u7684\u6cbb\u7642\u5e73\u53f0\u548c\u7528\u65bc\u6cbb\u7642\u6d3b\u6027\u751f\u7269\u88fd\u5291\u7684\u65b0\u578b\u905e\u9001\u7cfb\u7d71\u3002<\/p>\n

\"\"<\/p>\n","protected":false},"excerpt":{"rendered":"

\u5167\u76ae Transwell \u901a\u900f\u6027\u6aa2\u6e2c\u8a66\u5291\u76d2-Endothelial Transwell Permeabilit […]<\/p>\n","protected":false},"author":3,"featured_media":42664,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":[],"categories":[5628],"tags":[5776],"_links":{"self":[{"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/42663"}],"collection":[{"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/users\/3"}],"replies":[{"embeddable":true,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=42663"}],"version-history":[{"count":2,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/42663\/revisions"}],"predecessor-version":[{"id":42667,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/42663\/revisions\/42667"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/media\/42664"}],"wp:attachment":[{"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=42663"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=42663"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=42663"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}