{"id":40444,"date":"2023-03-23T17:51:23","date_gmt":"2023-03-23T09:51:23","guid":{"rendered":"http:\/\/biopioneer.com.tw\/?p=40444"},"modified":"2023-08-07T16:05:32","modified_gmt":"2023-08-07T08:05:32","slug":"7-aad-7-aminoactinomycin-d-%e8%9e%a2%e5%85%89-dna-%e7%b5%90%e5%90%88%e6%9f%93%e6%96%99-biotium-%e8%b2%a8%e8%99%9f40084-1mgml","status":"publish","type":"post","link":"http:\/\/biopioneer.com.tw\/?p=40444","title":{"rendered":"7-AAD (7-aminoactinomycin D) \u87a2\u5149 DNA \u7d50\u5408\u67d3\u6599 | Biotium \u8ca8\u865f40084 (1mg\/ml)"},"content":{"rendered":"<p><strong><span style=\"color: #33cccc; font-size: 12pt;\"><a style=\"color: #33cccc;\" href=\"http:\/\/biopioneer.com.tw\/?p=40444\">7-AAD (7-aminoactinomycin D) \u87a2\u5149 DNA \u7d50\u5408\u67d3\u6599 | Biotium \u8ca8\u865f40084<\/a>\u00a0(1mg\/ml)\u00a0<\/span><\/strong><\/p>\n<p>7-AAD\uff087-\u6c28\u57fa\u653e\u7dda\u83cc\u7d20 D, 7-aminoactinomycin D\uff09\u662f\u4e00\u7a2e\u819c\u4e0d\u53ef\u6ef2\u900f(membrane-impermeant )\u7684\u87a2\u5149 DNA \u7d50\u5408\u67d3\u6599\uff0c\u53ef\u7528\u65bc\u901a\u904e\u6d41\u5f0f\u7d30\u80de\u5100(\u00a0flow cytometry) \u9032\u884c\u6d3b\/\u6b7b\u7d30\u80de\u5340\u5206(\u00a0live\/dead discrimination )\u548c\u7d30\u80de\u9031\u671f\u5206\u6790(cell cycle profiling)\u30027-AAD \u662f\u4e00\u7a2e\u87a2\u5149 DNA \u7d50\u5408\u67d3\u6599(fluorescent DNA binding dye )\uff0c\u4e0d\u900f\u819c(membrane impermeant )\uff0c\u56e0\u6b64\u901a\u5e38\u88ab\u6d3b\u7d30\u80de(\u00a0live cells )\u548c\u65e9\u671f\u51cb\u4ea1\u7d30\u80de(\u00a0early apoptotic cells)\u6392\u9664\uff0c\u4f46\u6703\u67d3\u8272\u58de\u6b7b(\u00a0necrotic )\u548c\u665a\u671f\u51cb\u4ea1\u7d30\u80de(late apoptotic cells )\uff0c\u819c\u5b8c\u6574\u6027\u53d7\u640d(\u00a0membrane integrity)\u3002<\/p>\n<p><img decoding=\"async\" src=\"http:\/\/biopioneer.com.tw\/wp-content\/uploads\/2023\/03\/BIOTIUM_%E5%B7%A5%E4%BD%9C%E5%8D%80%E5%9F%9F-1-300x300.jpg\" alt=\"BIOTIUM_\u5de5\u4f5c\u5340\u57df 1\" \/><\/p>\n<p>&nbsp;<\/p>\n<p>\u8207 DNA \u7d50\u5408\u7684 7-AAD \u5728\u6fc0\u767c\/\u767c\u5c04\u6ce2\u9577\u70ba 546\/647 nm \u6642\u5177\u6709\u8f03\u5927\u7684Stokes shift\u00a0\u3002\u67d3\u6599\u53ef\u4ee5\u7528 488 nm \u6216 532 nm \u6fc0\u5149\u7dda\u6fc0\u767c\uff0c\u4e26\u5728 PE-Cy\u00ae5\/PerCP \u6d41\u5f0f\u7d30\u80de\u5100\u901a\u9053\u4e2d\u6aa2\u6e2c\u3002 7-AAD \u5ee3\u6cdb\u7528\u65bc\u5340\u5206\u6d3b\u7d30\u80de\/\u6b7b\u7d30\u80de\u3002\u5b83\u9084\u53ef\u7528\u65bc\u56fa\u5b9a\u548c\u900f\u5316\u7d30\u80de\uff0c\u901a\u904e\u4f7f\u7528\u6d41\u5f0f\u7d30\u80de\u5100\u9032\u884c DNA \u542b\u91cf\u5206\u6790\u4f86\u9032\u884c\u7d30\u80de\u9031\u671f\u5206\u6790\u3002 7-AAD \u9078\u64c7\u6027\u5730\u5d4c\u5165\u5bcc\u542b GC \u7684 DNA \u5340\u57df\uff0c\u4f7f\u5176\u67d3\u8272\u53ef\u7528\u65bc\u67d3\u8272\u9ad4\u986f\u5e36\u7814\u7a76\u3002\u6211\u5011\u63d0\u4f9b\u56fa\u9ad4\u7c89\u672b\u5f62\u5f0f\u7684 7-AAD\uff08\u8ca8\u865f\u00a040037\uff09\u6216 1 mg\/mL DMSO: \u6c34 (1:1) \u6eb6\u6db2\uff08\u8ca8\u865f40084\uff09\u3002\u5efa\u8b70\u4f7f\u7528 1 ug\/mL\uff081:1000 \u7a00\u91cb\uff09\u7684\u67d3\u6599\u4f86\u6aa2\u6e2c\u6d3b\/\u6b7b\u7d30\u80de\u6216 10 ug\/mL\uff081:100 \u7a00\u91cb\uff09\u7528\u65bc\u7d30\u80de\u9031\u671f\u5206\u6790\u3002<\/p>\n<p><img decoding=\"async\" loading=\"lazy\" class=\"\" src=\"http:\/\/biopioneer.com.tw\/wp-content\/uploads\/downloads\/2023\/03\/2-7-AAD-7-aminoactinomycin-D-%E8%9E%A2%E5%85%89-DNA-%E7%B5%90%E5%90%88%E6%9F%93%E6%96%99-Biotium-%E8%B2%A8%E8%99%9F40084.jpg\" alt=\"\" width=\"391\" height=\"286\" \/><\/p>\n<p><img decoding=\"async\" src=\"http:\/\/biopioneer.com.tw\/wp-content\/uploads\/downloads\/2023\/03\/BIOTIUM-40084.png\" alt=\"\" \/><\/p>\n<p><span style=\"color: #800000;\"><strong><span style=\"font-size: 12pt;\">7-AAD (7-aminoactinomycin D) is a membrane-impermeant fluorescent DNA binding dye that is useful for live\/dead discrimination and cell cycle profiling by flow cytometry.<\/span><\/strong><\/span><\/p>\n<ul>\n<li>Selective detection of dead cells by flow cytometry in the PE-Cy\u00ae5\/PerCP channel<\/li>\n<li>Perform cell cycle profiling by flow cytometry in fixed\/permeabilized cells<\/li>\n<li>\u03bb<sub>Ex<\/sub>\/\u03bb<sub>Em<\/sub>\u00a0(with DNA) = 546\/647 nm (7-AAD can also be excited by the 488 or 532 laser lines)<\/li>\n<li>Orange red solid soluble in DMF or DMSO<\/li>\n<li>Store at -20\u00b0C and protect from light, especially in solution<\/li>\n<li>C<sub>62<\/sub>H<sub>87<\/sub>N<sub>13<\/sub>O<sub>16<\/sub><\/li>\n<li>MW: 1270.45<\/li>\n<\/ul>\n<p><span style=\"color: #800000;\"><strong><span style=\"font-size: 12pt;\">\u5efa\u8b70\u4f7f\u7528<\/span><\/strong><\/span><\/p>\n<p>\u5efa\u8b70\u4f7f\u7528 1 ug\/mL\uff081:1000 \u7a00\u91cb\uff09\u7684\u67d3\u6599\u4f86\u6aa2\u6e2c\u6d3b\/\u6b7b\u7d30\u80de\u6216 10 ug\/mL\uff081:100 \u7a00\u91cb\uff09\u7528\u65bc\u7d30\u80de\u9031\u671f\u5206\u6790<\/p>\n<p>We recommend using the dye at 1 ug\/mL (1:1000 dilution) for live\/dead discrimination or 10 ug\/mL (1:100 dilution) for cell cycle profiling. Example protocols are provided below.<\/p>\n<p>&nbsp;<\/p>\n<p><img decoding=\"async\" loading=\"lazy\" class=\"\" src=\"http:\/\/biopioneer.com.tw\/wp-content\/uploads\/downloads\/2023\/03\/3-7-AAD-7-aminoactinomycin-D-%E8%9E%A2%E5%85%89-DNA-%E7%B5%90%E5%90%88%E6%9F%93%E6%96%99-Biotium-%E8%B2%A8%E8%99%9F40084.jpg\" alt=\"\" width=\"382\" height=\"438\" \/><\/p>\n<p><img decoding=\"async\" loading=\"lazy\" class=\"\" src=\"http:\/\/biopioneer.com.tw\/wp-content\/uploads\/downloads\/2023\/03\/7-AAD-7-aminoactinomycin-D-%E8%9E%A2%E5%85%89-DNA-%E7%B5%90%E5%90%88%E6%9F%93%E6%96%99-Biotium-%E8%B2%A8%E8%99%9F40084.jpg\" alt=\"\" width=\"388\" height=\"410\" \/><\/p>\n<table id=\"tablepress-307\" class=\"tablepress tablepress-id-307\">\n<thead>\n<tr class=\"row-1\">\n<th class=\"column-1\">Product<\/th>\n<th class=\"column-2\">Catalog Number<\/th>\n<th class=\"column-3\">Unit Size<\/th>\n<th class=\"column-4\">Format<\/th>\n<\/tr>\n<\/thead>\n<tbody class=\"row-hover\">\n<tr class=\"row-2\">\n<td class=\"column-1\">7-AAD<\/td>\n<td class=\"column-2\">40037<\/td>\n<td class=\"column-3\">1 mg<\/td>\n<td class=\"column-4\">Orange\/red solid<\/td>\n<\/tr>\n<tr class=\"row-3\">\n<td class=\"column-1\">7-AAD Solution, 1 mg\/mL<\/td>\n<td class=\"column-2\">40084<\/td>\n<td class=\"column-3\">1 mL<\/td>\n<td class=\"column-4\">Red solution in DMSO:Water (1:1)<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p><span style=\"color: #800000;\"><strong><span style=\"font-size: 12pt;\">Experimental Protocols <\/span><\/strong><\/span><\/p>\n<p><span style=\"color: #339966;\">For live\/dead discrimination by flow cytometry <\/span><\/p>\n<p>1. Prepare a positive control by incubating cells at 56\u00b0C for 30 minutes then cool to room temperature. Include an untreated cell sample as a negative control.<\/p>\n<p>2. Adjust cells to 5&#215;106 cells per mL in complete culture medium or buffer of your choice and aliquot 1 mL per flow tube. Note: Cells can be stained anywhere between 5&#215;105 cells\/mL to 10<sup>7<\/sup> cells per mL in 100 uL to 1 mL. If necessary, a 100 ug\/mL intermediate dilution of 7-AAD can be prepared by diluting the stock solution 1:10 in water or buffer.<\/p>\n<p>3. Add 1 uL of 1 mg\/mL 7-AAD to 1 mL of cells and mix.<\/p>\n<p>4. Incubate 15-30 minutes at room temperature, protected from light. The incubation can be carried out on ice if desired.<\/p>\n<p>5. Analyze by flow cytometry in the PE-Cy\u00ae5 or PerCP channel without washing the cells.<\/p>\n<p>Notes: a. While 7-AAD staining is retained after formaldehyde fixation, separation between live and dead cells is reduced after fixation due to dye transfer from dead to live cells. For truly fixable dead cell staining, we recommend using a covalent dye such as our Live-or-Dye\u2122 stains (see Related Products). b. This protocol was optimized using Jurkat cells. Assay optimization may be required for use with other cell types. c. If you prefer not to wash your cells, staining can be performed in cell culture medium with serum instead of buffer.<\/p>\n<p><span style=\"color: #800000;\"><strong><span style=\"font-size: 12pt;\">For cell cycle profiling by flow cytometry analysis of DNA content <\/span><\/strong><\/span><\/p>\n<p><span style=\"color: #339966;\">Materials required but not provided (see Related Products)<\/span><\/p>\n<p>\u2022 Flow Cytometry Fixation\/Permeabilization Kit (catalog no. 23006)<\/p>\n<p>\u2022 1X Phosphate buffered saline (PBS) or your preferred FACS buffer Staining Protocol<\/p>\n<p>1. Adjust cells to 10<sup>7<\/sup> cells per mL and aliquot 100 uL per flow tube.<\/p>\n<p>2. Fix and permeabilize cells according the protocol for the Flow Cytometry Fixation\/Permeabilization Kit, or use your preferred method.<\/p>\n<p>3. Pellet the cells by centrifugation and wash with 1X PBS or FACS buffer.<\/p>\n<p>4. Pellet the cells by centrifugation and resuspend in 100 uL buffer.<\/p>\n<p>5. Add 1 uL of 1 mg\/mL 7-AAD per tube and mix by gentle vortexing.<\/p>\n<p>6. Incubate 15 minutes at room temperature, protected from light.<\/p>\n<p>7. Add 400 uL PBS or FACS buffer per tube. Analyze by flow cytometry in the PE-Cy\u00ae5 channel or PerCP channel. Use a linear scale for fluorescence detection, and acquire data with a slow flow rate (~12 uL \/minute).<\/p>\n<p><a href=\" http:\/\/biopioneer.com.tw\/wp-content\/uploads\/downloads\/2023\/03\/7-AAD-7-aminoactinomycin-D-\u87a2\u5149-DNA-\u7d50\u5408\u67d3\u6599-Biotium-\u8ca8\u865f40084.pdf\"><img decoding=\"async\" loading=\"lazy\" class=\"alignnone\" src=\"http:\/\/biopioneer.com.tw\/wp-content\/uploads\/2023\/01\/%E7%B6%B2%E7%AB%99%E8%A6%8F%E6%A0%BC%E5%B0%8F%E5%9C%96_%E5%B7%A5%E4%BD%9C%E5%8D%80%E5%9F%9F-1_%E5%B7%A5%E4%BD%9C%E5%8D%80%E5%9F%9F-11-300x97.jpg\" alt=\"\u7db2\u7ad9\u898f\u683c\u5c0f\u5716_\u5de5\u4f5c\u5340\u57df 1_\u5de5\u4f5c\u5340\u57df 1\" width=\"149\" height=\"48\" \/><\/a><\/p>\n<p>1. Environ Mol Mutagen (2013) 54(8), 682-9.\u00a0<a href=\"https:\/\/onlinelibrary.wiley.com\/doi\/abs\/10.1002\/em.21800\" target=\"_blank\" rel=\"noopener noreferrer\">doi: 10.1002\/em.21800<\/a><br \/>\n2. Mol Cell Biol (2014) 34(24), 4485\u20134499.\u00a0<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4248725\/\" target=\"_blank\" rel=\"noopener noreferrer\">doi: 10.1128\/MCB.00671-14<\/a><br \/>\n3. Mutat Res Genet Toxicol Environ Mutagen (2014) 770, 1-5.\u00a0<a href=\"https:\/\/www.sciencedirect.com\/science\/article\/abs\/pii\/S138357181400134X?via%3Dihub\" target=\"_blank\" rel=\"noopener noreferrer\">doi: 10.1016\/j.mrgentox.2014.05.001<\/a><br \/>\n4. Science (2017) 356(6339) eaai9264.\u00a0<a href=\"https:\/\/science.sciencemag.org\/content\/356\/6339\/eaai9264\" target=\"_blank\" rel=\"noopener noreferrer\">DOI: 10.1126\/science.aai9264<\/a><br \/>\n5. Int J Rad Biol (2019) 95,9, 1226-1235.\u00a0<a href=\"https:\/\/www.tandfonline.com\/doi\/abs\/10.1080\/09553002.2019.1625490\" target=\"_blank\" rel=\"noopener noreferrer\">DOI: 10.1080\/09553002.2019.1625490<\/a><br \/>\n6. OncoTargets Therapy (2019) 12, 5227-5239.\u00a0<a href=\"https:\/\/www.dovepress.com\/ethyl-acetate-leave-extract-of-nepenthes-ventricosa-x-sibuyanensis-cau-peer-reviewed-article-OTT\" target=\"_blank\" rel=\"noopener noreferrer\">https:\/\/doi.org\/10.2147\/OTT.S190460<\/a><br \/>\n7. Antioxidants (2020) 9(9), 873.\u00a0<a href=\"https:\/\/www.mdpi.com\/2076-3921\/9\/9\/873876\" target=\"_blank\" rel=\"noopener noreferrer\">doi:10.3390\/antiox9090873<\/a><br \/>\n8. Antioxidants (2020) 9(9), 876.\u00a0<a href=\"https:\/\/www.mdpi.com\/2076-3921\/9\/9\/876\" target=\"_blank\" rel=\"noopener noreferrer\">doi:10.3390\/antiox9090876<\/a><\/p>\n<p><img decoding=\"async\" loading=\"lazy\" class=\"\" src=\"http:\/\/biopioneer.com.tw\/wp-content\/uploads\/2023\/03\/%E5%93%81%E7%89%8C%E4%BB%8B%E7%B4%B9_%E5%B7%A5%E4%BD%9C%E5%8D%80%E5%9F%9F-1-300x120.jpg\" alt=\"\u54c1\u724c\u4ecb\u7d39_\u5de5\u4f5c\u5340\u57df 1\" width=\"146\" height=\"58\" \/><\/p>\n<p><img decoding=\"async\" loading=\"lazy\" class=\"\" src=\"http:\/\/biopioneer.com.tw\/wp-content\/uploads\/2023\/03\/biotium-300x116.jpg\" alt=\"biotium\" width=\"226\" height=\"75\" \/><\/p>\n<p><strong><span style=\"font-size: 12pt; color: #800000;\">Biotium \u63d0\u4f9b\u7bc4\u570d\u5ee3\u6cdb\u7684\u87a2\u5149\u63a2\u91dd\u7528\u65bc\u7814\u7a76\u51cb\u4ea1\u7d30\u80de(Apotosis)<\/span><\/strong><\/p>\n<p><img decoding=\"async\" src=\"http:\/\/biopioneer.com.tw\/wp-content\/uploads\/downloads\/2023\/03\/biotium.jpg\" alt=\"\" \/><\/p>\n<p>The\u00a0<a href=\"https:\/\/biotium.com\/product\/nucview-caspase-3-substrates\/\" target=\"_blank\" rel=\"noopener noreferrer\">NucView\u00ae Caspase-3 Substrate<\/a>\u00a0comes in 3 colors, and measures caspase activity in real time, in live cells.\u00a0<a href=\"https:\/\/biotium.com\/product\/mitoview-dyes\/\" target=\"_blank\" rel=\"noopener noreferrer\">MitoView\u2122 633<\/a>\u00a0can measure the decrease in mitochondria membrane potential that occurs in early apoptosis.\u00a0<a href=\"https:\/\/biotium.com\/product\/annexin-v-conjugates\/\" target=\"_blank\" rel=\"noopener noreferrer\">CF\u00ae Dye Annexin V<\/a>\u00a0conjugates come in many color options, and label PS on the membranes of late apoptotic cells. We also offer kits containing combinations of these probes.<\/p>\n","protected":false},"excerpt":{"rendered":"<p>7-AAD (7-aminoactinomycin D) \u87a2\u5149 DNA \u7d50\u5408\u67d3\u6599 | Biotium \u8ca8\u865f40 [&hellip;]<\/p>\n","protected":false},"author":3,"featured_media":40136,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":[],"categories":[5378],"tags":[5407],"_links":{"self":[{"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/40444"}],"collection":[{"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/users\/3"}],"replies":[{"embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=40444"}],"version-history":[{"count":4,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/40444\/revisions"}],"predecessor-version":[{"id":44891,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/40444\/revisions\/44891"}],"wp:featuredmedia":[{"embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/media\/40136"}],"wp:attachment":[{"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=40444"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=40444"},{"taxonomy":"post_tag","embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=40444"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}