{"id":44565,"date":"2023-07-28T17:10:11","date_gmt":"2023-07-28T09:10:11","guid":{"rendered":"https:\/\/biopioneer.com.tw\/?p=44565"},"modified":"2023-08-30T11:33:13","modified_gmt":"2023-08-30T03:33:13","slug":"20s%e8%9b%8b%e7%99%bd%e9%85%b6%e9%ab%94%e6%b8%ac%e5%ae%9a%e8%a9%a6%e5%8a%91%e7%9b%92-20s-proteasome-assay-kit-%e8%b2%a8%e8%99%9fabnova-ka1352","status":"publish","type":"post","link":"http:\/\/biopioneer.com.tw\/?p=44565","title":{"rendered":"20S\u86cb\u767d\u9176\u9ad4\u6e2c\u5b9a\u8a66\u5291\u76d2| 20S Proteasome Assay Kit \u8ca8\u865fAbnova\u2122 KA1352"},"content":{"rendered":"

20S\u86cb\u767d\u9176\u9ad4\u6e2c\u5b9a\u8a66\u5291\u76d2| 20S Proteasome Assay Kit \u8ca8\u865fAbnova\u2122 KA1352<\/span><\/a><\/strong><\/span><\/p>\n

<\/p>\n

\u86cb\u767d\u9176\u9ad4(proteasome)\u662f\u4e00\u7a2e\u591a\u50ac\u5316\u86cb\u767d\u9176\u8907\u5408\u7269(multicatalytic proteinase complex )\uff0c\u53c3\u8207\u9078\u64c7\u6027\u964d\u89e3\u7d30\u80de\u5167\u86cb\u767d\u8cea\u3002 20S\u86cb\u767d\u9176\u9ad4( 20S proteasome)\u662f\u5927\u86cb\u767d\u964d\u89e3\u7684\u86cb\u767d\u6c34\u89e3\u6838\u5fc3\u9846\u7c92\u8907\u96dc\u768426S\u86cb\u767d\u9176\u9ad4( 26S proteasome)\u3002\u9019\u7a2e 700 kDa \u7684\u86cb\u767d\u8cea\u666e\u904d\u5206\u4f48\u5728\u7d30\u80de\u8cea\u548c\u7d30\u80de\u8cea\u4e2d\u3002\u6240\u6709\u771f\u6838\u751f\u7269\u7684\u7d30\u80de\u6838\u3002 \u8a72\u86cb\u767d\u8cea\u8ca0\u8cac\u964d\u89e3\u8a31\u591a\u7d30\u80de\u9031\u671f\u63a7\u5236\u56e0\u5b50(cell-cycle control factors)\u3001\u4fe1\u865f\u8f49\u5c0e\u56e0\u5b50(signal transduction factors)\u3001\u8f49\u9304\u56e0\u5b50( transcription factors)\u4ee5\u53ca\u764c\u57fa\u56e0(oncogene)\u548c\u6297\u764c\u57fa\u56e0\u7522\u7269(anti-oncogene products)\uff0c\u5f9e\u800c\u63a7\u5236\u7d30\u80de\u589e\u6b96(cell proliferation)\u3001\u5206\u5316( differentiation)\u548c\u51cb\u4ea1(apoptosis)\u3002 20S\u86cb\u767d\u9176\u9ad4( 20S proteasome)\u8868\u9054\u7684\u8b8a\u5316\u8a18\u9304\u5728\u8a31\u591a\u751f\u7406\u548c\u75c5\u7406\u689d\u4ef6\u4e0b\uff0c\u4f8b\u5982\u9aa8\u9abc\u808c\u840e\u7e2e(skeletal muscle atrophy )\u548c\u963f\u723e\u8328\u6d77\u9ed8\u795e\u7d93\u8b8a\u6027(Alzheimer neurodegeneration)\u3002\u4e59\u9187\u6291\u5236\u86cb\u767d\u9176\u9ad4\u6d3b\u6027(Ethanol inhibits proteasome activity)\uff0c\u5c0e\u81f4\u4e0d\u6eb6\u6027\u7269\u8cea\u7a4d\u7d2f\u88ab\u7a31\u70ba\u99ac\u6d1b\u91cc\u9ad4\u7684\u86cb\u767d\u8cea\u805a\u96c6\u9ad4\uff0c\u662f\u9152\u7cbe\u6027\u809d\u75c5(alcoholic liver diseases)\u7684\u5e38\u898b\u7279\u5fb5\u3002 \u53e6\u4e00\u65b9\u9762\uff0c\u86cb\u767d\u9176\u9ad4\u6291\u5236\u5291\u8868\u73fe\u51fa\u6297\u708e(anti-inflammatory )\u548c\u6297\u589e\u6b96\u4f5c\u7528(antiproliferative effects)\uff0c\u8b49\u64da\u8868\u660e\u86cb\u767d\u9176\u9ad4\u53ef\u80fd\u662f\u6cbb\u7642\u764c\u75c7\u548c\u708e\u75c7\u6027\u75be\u75c5\u7684\u91cd\u8981\u85e5\u7269\u9776\u9ede\u3002<\/p>\n

\u6e2c\u5b9a\u539f\u7406 (Principle of the Assay)<\/span><\/strong><\/span><\/p>\n

20S \u86cb\u767d\u9176\u9ad4\u6aa2\u6e2c\u8a66\u5291\u76d2(The 20S Proteasome Assay Kit) \u63a1\u7528\u7279\u5b9a\u7684 20S \u5e95\u7269 SUC-LLVY-AMC\uff0c\u5728\u88c2\u89e3\u5f8c\u6d3b\u6027\u9176\u7522\u751f\u9ad8\u87a2\u5149\u7522\u7269\uff0c\u53ef\u4ee5\u4f7f\u7528\u6fc0\u767c\u548c\u767c\u5c04\u9032\u884c\u6e2c\u91cf\u6ce2\u9577\u5206\u5225\u70ba360 nm\u548c480 nm\u3002\u8a72\u5957\u4ef6\u6613\u65bc\u4f7f\u7528\uff0c\u53ef\u8f15\u9b06\u9069\u61c9\u9ad8\u8abf\u7bc0 20S \u86cb\u767d\u9176\u9ad4\u6fc0\u6d3b\u7684\u6cbb\u7642\u5316\u5408\u7269\u7684\u901a\u91cf\u7be9\u9078\u3002jurkat cell lysate supernatant\u5305\u542b\u542b\u6709\u9ad8\u6c34\u5e73 20S \u6d3b\u6027\u7684\u88c2\u89e3\u7269\u4e0a\u6e05\u6db2\uff0c\u7528\u4f5c\u967d\u6027\u5c0d\u7167\u3002\u8a66\u5291\u76d2\u4e2d\u9084\u5305\u542b\u7528\u65bc\u8b49\u660e\u7279\u7570\u6027\u7684\u7684\u57fa\u6750\u4e00\u7a2e\u7279\u5b9a\u7684 20S \u6291\u88fd\u5291\u8868\u6c92\u98df\u5b50\u5152\u8336\u7d20\u6c92\u98df\u5b50\u9178\u916f (epigallocatechin gallate, EGCG)\u3002<\/p>\n

<\/p>\n

\"\u7db2\u7ad9\u898f\u683c\u5c0f\u5716_\u5de5\u4f5c\u5340\u57df<\/a><\/p>\n

20S Proteasome Activity in Jurkat Cell Lysate Supernatant.<\/p>\n

<\/p>\n

Figure 1: 20S Proteasome Activity in Jurkat Cell Lysate Supernatant. Jurkat cells were seeded in a 96-well plate in 100 \u03bcL of culture medium at a density indicated on the graph.
\nCells were then processed for measurement of 20S activity according to the protocol described in the Assay Procedure section. Relative fluorescent intensity was then measured with a plate reader.
\nNote: Lactacystin is a known selective 20S inhibitor.<\/p>\n

 <\/p>\n

<\/p>\n

Figure 3: H2O2 inhibits 20S Proteasome Activity in Raw 264.7 cells<\/p>\n

Raw 264.7 cells were seeded in a 96-well plate in 100 \u03bcL of culture medium at 8 x 104 cells\/well and grown in DMEM containing 10% FBS for three days. Cells were then treated with different doses of H2O2 as indicated on the chart for 30 minutes and processed for measurement of 20S activity according to the protocol described in the Performing the Assay section. Relative fluorescent intensity was then measured with a plate reader.<\/p>\n

References<\/span><\/strong><\/span>
\n1. Zwickl, P., Voges, D., and Baumeister, W. The proteasome: a macromolecular assembly designed for
\ncontrolled proteolysis. Philos Trans R Soc Lond B Biol Sci. 354, 1501-1511 (1999).
\n2. DeMartino, G.N. and Slaughter, C.A. The proteasome, a novel protease regulated by multiple mechanisms.
\nJ. Biol. Chem. 274(32), 22123-22126 (1999).
\n3. Osna, N.A. and Donohue, T.M., Jr. Implication of altered proteasome function in alcoholic liver injury.
\nWorld J. Gastroenterol 13(37), 4931-4937 (2007).
\n4. Elliott, P.J., Zollner, T.M., and Boehncke, W.-H. Proteasome inhibition: A new anti-inflammatory strategy. J.
\nMol. Med. 81, 235-245 (2003).
\n5. Smith, D.M., Wang, Z., Kazi, A., et al. Synthetic analogs of green tea polyphenols as proteasome inhibitors.
\nMol. Med. 8(7), 382-392 (2002).<\/p>\n","protected":false},"excerpt":{"rendered":"

20S\u86cb\u767d\u9176\u9ad4\u6e2c\u5b9a\u8a66\u5291\u76d2| 20S Proteasome Assay Kit \u8ca8\u865fAbnova\u2122 KA1352 […]<\/p>\n","protected":false},"author":3,"featured_media":40950,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":[],"categories":[5549],"tags":[6082],"_links":{"self":[{"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/44565"}],"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=44565"}],"version-history":[{"count":3,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/44565\/revisions"}],"predecessor-version":[{"id":45451,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/posts\/44565\/revisions\/45451"}],"wp:featuredmedia":[{"embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=\/wp\/v2\/media\/40950"}],"wp:attachment":[{"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=44565"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=44565"},{"taxonomy":"post_tag","embeddable":true,"href":"http:\/\/biopioneer.com.tw\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=44565"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}