{"id":7531,"date":"2016-11-10T12:47:10","date_gmt":"2016-11-10T04:47:10","guid":{"rendered":"http:\/\/biopioneer.com.tw\/?p=7531"},"modified":"2023-06-29T18:24:53","modified_gmt":"2023-06-29T10:24:53","slug":"%e8%84%82%e8%82%aa%e7%b5%84%e7%b9%94%e8%9b%8b%e7%99%bd%e8%b3%aa%e8%90%83%e5%8f%96%e5%a5%97%e7%b5%84-invent%e8%b2%a8%e8%99%9fat-022","status":"publish","type":"post","link":"http:\/\/biopioneer.com.tw\/?p=7531","title":{"rendered":"\u8102\u80aa\u7d44\u7e54\u86cb\u767d\u8cea\u8403\u53d6\u5957\u7d44 | Adipose Tissues\/Cultured Adipocyte Adipose Tissues\/Cultured Adipocyte Invent\u8ca8\u865fAT-022"},"content":{"rendered":"

\u8102\u80aa\u7d44\u7e54<\/strong>\/<\/strong>\u57f9\u990a\u4e4b\u8102\u80aa\u7d30\u80de\u86cb\u767d\u8cea<\/strong>\u8403\u53d6\u5957\u7d44<\/strong> |Invent<\/strong>\u8ca8\u865f<\/strong>AT-022<\/strong><\/a><\/span><\/p>\n

\u7522\u54c1\u540d\u7a31<\/strong>:\u00a0Minute\u2122 \u00a0<\/strong>\u8102\u80aa\u7d44\u7e54<\/strong>\/<\/strong>\u57f9\u990a\u4e4b\u8102\u80aa\u7d30\u80de\u86cb\u767d\u8cea<\/strong>\u8403\u53d6\u5957\u7d44<\/strong>| Adipose Tissues\/Cultured Adipocyte<\/strong>\u00a0for Adipose Tissues\/Cultured Adipocytes| \u8ca8\u865fAT-022<\/p>\n

\"\"<\/p>\n

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

\u7522\u54c1\u4ecb\u7d39:<\/strong> <\/span>\u8102\u80aa\u7d44\u7e54\u7279\u5225\u662f\u767d\u8272\u8102\u80aa\u7d44\u7e54\uff08WAT\uff09\u5df2\u7d93\u88ab\u8a8d\u70ba\u662f\u9664\u4e86\u5176\u80fd\u91cf\u5b58\u5132\u529f\u80fd\u4e4b\u5916\u7684\u91cd\u8981\u7684\u5167\u5206\u6ccc\u548c\u708e\u75c7\u5668\u5b98\u3002\u4f86\u81ea\u8102\u80aa\u7d44\u7e54\u7684\u86cb\u767d\u8cea\u7684\u5206\u96e2\u8207\u5206\u6790\u5c0d\u65bc\u7406\u89e3\u8a31\u591a\u751f\u7406\/\u75c5\u7406\u72c0\u6cc1\u8d8a\u4f86\u8d8a\u91cd\u8981\u3002\u7136\u800c\uff0c\u7531\u65bc\u5176\u9ad8\u8102\u8cea\u548c\u4f4e\u86cb\u767d\u8cea\u542b\u91cf\uff0c\u8b93WAT\u548c\u68d5\u8272\u8102\u80aa\u7d44\u7e54\uff08BAT\uff09\u7684\u5206\u96e2\u548c\u5206\u6790\u5728\u6280\u8853\u4e0a\u662f\u975e\u5e38\u5177\u6709\u6311\u6230\u6027\u7684\u3002\u5b58\u5728\u65bc\u751f\u7269\u6a23\u54c1\u4e2d\u7684\u6c34 – \u6cb9\u4e73\u6db2\u96e3\u4ee5\u5206\u96e2\uff1bInvent\u958b\u767c\u65b0\u6280\u8853\uff0c\u5177\u6709\u7368\u7279\u7684\u8868\u9762\u6027\u8cea\u548c\u9810\u5b9a\u7684\u5b54\u5f91\u548c\u539a\u5ea6\u7684\u591a\u5b54\u904e\u6ffe\u5668\u8207\u7279\u5225\u914d\u88fd\u7684\u4e0d\u542b\u6d17\u6ecc\u5291\u7684\u63d0\u53d6\u7de9\u885d\u6db2\u7d50\u5408\uff0c\u7528\u65bc\u5feb\u901f\u548c\u6709\u6548\u5730\u5206\u96e2\u5f97\u81ea\u8102\u80aa\u7d44\u7e54\u52fb\u6f3f\u7684\u6c34 – \u6cb9\u4e73\u6db2\u3002\u63d0\u53d6\u7de9\u885d\u6db2\u5177\u6709\u6bd4\u8102\u80aa\u7d44\u7e54\u4e2d\u7684\u6cb9\u66f4\u4f4e\u7684\u51dd\u56fa\u9ede\uff0c\u4e26\u4e14\u6c34\u76f8\u53ef\u4ee5\u901a\u904e\u4f7f\u7d44\u7e54\u52fb\u6f3f\u901a\u904e\u904e\u6ffe\u5668\u800c\u5f9e\u6cb9\u76f8\u5feb\u901f\u5206\u96e2\u3002\u8207\u5176\u4ed6\u65b9\u6cd5\u76f8\u6bd4\uff0c\u63d0\u53d6\u9ad8\u6fc3\u5ea6\u86cb\u767d\u8cea\uff082-3mg \/ ml\uff09\u542b\u91cf\u3002\u53ef\u61c9\u7528\u65bc2D gel analysis\u3001 ELISA\u3001 SDS-PAGE\u3001, immunoblotting, immunoprecipitation\u3001 enzyme activity assays\u3001\u7b49\u5206\u6790\u3002<\/p>\n

Minute\u2122<\/strong>\u86cb\u767d\u8403\u53d6\u5957\u7d44\uff0c\u4f7f\u7528\u5275\u65b0\u300e<\/strong>Column Filter<\/strong>\u300f\u6280\u8853\uff0c\u6548\u679c\u66f4\u4f73\uff0c\u5b8c\u6210\u53ea\u9700<\/strong>\u00a0<\/strong>\u4e00<\/strong> \u5206\u9418\u86cb\u767d\u8403\u53d6\u53ea\u9700\u8981<\/strong>vortex<\/em><\/strong>\u3001<\/em><\/strong>transfer<\/em><\/strong>\u3001<\/em><\/strong>spin <\/em><\/strong>\u3002<\/em><\/strong>Minute \u2122\u4e00\u7cfb\u5217\u86cb\u767d\u8cea\u63d0\u53d6\u8a66\u5291\u76d2\uff1b\u9069\u7528\u65bc\u591a\u7a2e\u7269\u7a2e\uff08\u5305\u62ec\u54fa\u4e73\u52d5\u7269\u3001\u6d77\u6d0b\u52d5\u7269\u3001\u6606\u87f2\u3001\u690d\u7269\u548c\u7d30\u83cc\uff09\u5feb\u901f\u548c\u5bb9\u6613\u5730\u63d0\u53d6\/\u5206\u96e2\u86cb\u767d\u8cea\u7684\u512a\u7570\u5de5\u5177 \u3002\u8207\u5e02\u5834\u4e0a\u7684\u4efb\u4f55\u5176\u4ed6\u86cb\u767d\u8cea\u63d0\u53d6\u8a66\u5291\u76d2\u4e0d\u540c\uff0c\u512a\u5316\u7684\u7de9\u885d\u6db2\u7cfb\u7d71\uff0c\u7d50\u5408\u6700\u65b0\u7684\u5c08\u6709\u6ffe\u7b52\u6280\u8853\uff08spin column\uff09\uff1b\u5feb\u901f\u63d0\u9ad8\u86cb\u767d\u8403\u53d6\u7684\u6548\u80fd\u53ca\u65b9\u4fbf\u6027\uff1b\u7528\u65bc\u7e3d\u86cb\u767d\u63d0\u53d6\uff0c\u7d30\u80de\u5206\u96e2\u548c\u819c\u86cb\u767d\u5206\u96e2\uff0c\u53ea\u9700\u7c21\u55ae\u56db\u6b65\u9a5f\u5373\u53ef\u5b8c\u6210\u86cb\u767d\u8cea\u8403\u53d6\u3002\u7121\u9808\u9577\u6642\u9593\u7b49\u5f85\u3001\u514d\u9664\u91cd\u8907\u6027\u7684wash\u6b65\u9a5f\u3002<\/p>\n

\"\"<\/p>\n

SDS-PAGE (10%) profiles of total protein extracted from different adipose tissues. Lane 1, porcine WAT; Lane 2, chicken BAT; Lane 3, rat WAT. B. Western blottings of extracted proteins from rat WAT. Proteins were separated in 8-16% gradient SDS-PAGE and probed with following cellular protein marker antibodies:Anti-Na\/K ATPase alpha1, a plasma membrane marker(Upstate, clone 464.6), anti-lamin B1, a nuclear envelope marker (ab16048, abcam Cambridge, MA), anti-ubiquinol-cytochrome C reductase core protein (abcam, ab 96333) and GAPDH, a cytosolic marker (Sigma). The specific protein bands were visualized by a substrate Opti-4CN (Bio-RAD).<\/p>\n

\"AT-022-data-2\"<\/p>\n

\u6ce8\u610f\u4e8b\u9805:\u00a0<\/span><\/p>\n

\u86cb\u767d\u8cea\u6700\u4f73\u5316\uff0c\u53ef\u4f7f\u7528protease inhibitors\u86cb\u767d\u9176\u6291\u88fd\u5291\u3002\u5982\u679c\u4e0b\u6e38\u61c9\u7528\u662f\u9700\u8981\u6bd4\u8f03\u4e45\u6642\u9593\uff1b\u6216\u86cb\u767d\u8cea\u63d0\u53d6\u7269\u5132\u5b58\u9577\u6642\u9593\u624d\u4f7f\u7528\uff0c\u53ef\u65bc\u7de9\u885d\u6db2A\u4e2d\u52a0\u5165protease inhibitors\u86cb\u767d\u9176\u6291\u88fd\u5291\u4f7f\u7528\u3002\u86cb\u767d\u8cea\u6fc3\u5ea6\u5b9a\u91cf\u53ef\u61c9\u7528BCA\u65b9\u6cd5\u6e2c\u5b9a\u3002 \u65bc\u86cb\u767d\u8cea\u78f7\u9178\u5316\u7814\u7a76\uff0c\u5728\u4f7f\u7528\u524d\u52a0\u5165\u78f7\u9178\u9176\u6291\u88fd\u5291cocktails of phosphatase inhibitors\u653e\u7de9\u885d\u6db2A\u4e2d\u4f7f\u7528\u3002**\u5982\u679c\u7de9\u885d\u6db2B\u5728\u8f03\u4f4e\u6eab\u5ea6\u4e0b\u5b58\u5728\u6c89\u6fb1\u7269\uff0c37\u2103\u56de\u6eab\u76f4\u5230\u6c89\u6fb1\u5b8c\u5168\u6eb6\u89e3\u3002<\/p>\n

\u63d0\u53d6\u7684\u86cb\u767d\u8cea\u770b\u8d77\u4f86\u7565\u5fae\u6a21\u7cca\u4e0d\u6e05\u5b58\u5728\u6c34\u4e0d\u6eb6\u6027\u7d30\u80de\u6210\u5206\u3002\u5b83\u53ef\u4ee5\u7a00\u91cb\u4e26\u76f4\u63a5\u7528\u65bcELISA\u6aa2\u6e2c\u6c34\u6eb6\u6027\u86cb\u767d\u8cea\u3002\u5b83\u4e5f\u53ef\u4ee5\u91cd\u61f8\u65bc\u7de9\u885d\u6db2B ( buffer B) \u6216\u7de9\u885d\u6db2C ( buffer C)\u4e2d\uff0c\u4ee5\u6eb6\u89e3\u6c34\u4e0d\u6eb6\u6027\u86cb\u767d\u8cea (water insoluble proteins ) \u4e26\u7528\u65bc\u4e0b\u6e38\u61c9\u7528\uff1a
\nA.\u63d0\u53d6\u7684\u86cb\u767d\u8cea\u6eb6\u6db2\u4e2d\u52a0\u51651\/10\u7684\u7de9\u885d\u6db2B (\u00a0buffer B)\uff0c\u5f97\u5230\u8b8a\u6027\u7684\u86cb\u767d\u8cea\u6eb6\u6db2\uff08\u9069\u7528\u65bcSDS-PAGE\uff0cWesterns\u548c\u5176\u4ed6\u61c9\u7528\uff09
\nB.\u63d0\u53d6\u7684\u86cb\u767d\u8cea\u6eb6\u6db2\u4e2d\u52a0\u51651\/10\u7684\u7de9\u885d\u6db2C (\u00a0buffer B)\uff0c\u5f97\u5230\u672a\u8b8a\u6027\u7684\u86cb\u767d\u8cea\uff08\u9069\u7528\u65bcIP\uff0cELISA\u548c\u5176\u4ed6\u61c9\u7528\uff09
\nC.\u6eb6\u65bc2\u00d72D\u51dd\u81a0\u6a23\u54c1\u7de9\u885d\u6db2\u4e2d\u9032\u884c2D\u51dd\u81a0\u5206\u6790<\/p>\n

\u6ce8\u610f\uff1a\u7de9\u885d\u6db2A\u5305\u542b\u53ef\u80fd\u5e72\u64fe\u8cea\u8b5c\u5206\u6790\u7684\u7d44\u5206\u3002\u5982\u679c\u63d0\u53d6\u86cb\u767d\u8cea\u7528\u65bcMS\u5206\u6790\uff0c\u9996\u5148\u91dd\u5c0d\u517c\u5bb9\u7de9\u885d\u6db2\u9032\u884c\u900f\u6790\u3002\u53e6\u4e00\u7a2e\u9078\u64c7\u662f\u6c89\u6fb1\u86cb\u767d\u8cea\u3002\u4f7f\u7528\u6a19\u6e96TCA\u86cb\u767d\u6c89\u6fb1\u65b9\u6848\u3002<\/p>\n

\n\n\n\n\n\n\n\n\n\n\n
Item<\/td>\nAmount<\/td>\n<\/tr>\n
Buffer A (Extraction buffer)<\/td>\n15ml<\/td>\n<\/tr>\n
Buffer B (10 X denaturing buffer)<\/td>\n1.5ml<\/td>\n<\/tr>\n
Buffer C (10 X non-denaturing buffer)<\/td>\n1.5ml<\/td>\n<\/tr>\n
1.5 ml microfuge tube<\/td>\n20<\/td>\n<\/tr>\n
Pestles for 1.5 ml tube<\/td>\n2<\/td>\n<\/tr>\n
Filter Cartridge with Collection Tubes<\/td>\n20<\/td>\n<\/tr>\n
Protein Extraction Powder<\/td>\n2g<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<\/div>\n

\u5f15\u7528\u6587\u737b<\/span><\/strong><\/span><\/p>\n

MinuteTM<\/sup>\u00a0Total Protein Extraction Kit for Adipose Tissue\/Cultured cells (AT-022)<\/b><\/p>\n

Wang, N., Liu, Y., Ma, Y., & Wen, D. (2017). High-intensity interval versus moderate-intensity continuous training: Superior metabolic benefits in diet-induced obesity mice. Life Sciences.
\nZheng, Q., Lin, J., Huang, J., Zhang, H., Zhang, R., Zhang, X., … & Song, R. (2017). Reconstitution of UCP1 using CRISPR\/Cas9 in the white adipose tissue of pigs decreases fat deposition and improves thermogenic capacity. Proceedings of the National Academy of Sciences, 201707853.
\nInvent, B. I. (2017). High Efficiency Protein Extraction from Adipose Tissues. BioTechniques, 63(5), 235
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\nChen, G., Li, Q., Luo, Y., Liu, T., Zhou, S., Pan, E., & Peng, L. (2018). Effect of Notoginsenoside R1 on autologous adipose graft in rats. Molecular Medicine Reports.
\nChen, T. X., Cheng, X. Y., Wang, Y., & Yin, W. (2018). Toosendanin inhibits adipogenesis by activating Wnt\/\u03b2-catenin signaling. Scientific Reports, 8(1), 4626.
\nWang, N., Liu, Y., Ma, Y., & Wen, D. (2018). Hydroxytyrosol ameliorates insulin resistance by modulating endoplasmic reticulum stress and prevents hepatic steatosis in diet-induced obesity mice. The Journal of Nutritional Biochemistry.
\nDu, W., Piek, A., Schouten, E. M., van de Kolk, C. W., Mueller, C., Mebazaa, A., … & Sillj\u00e9, H. H. Plasma levels of heart failure biomarkers are primarily a reflection of extracardiac production. Theranostics, Vol. 8 4155-4169
\nHann, M. (2018). Anti-inflammatory Activity of Isomaltodextrin in a C57BL\/6NCrl Mouse Model with Lipopolysaccharide-induced Low-grade Chronic Inflammation (Doctoral dissertation).
\nLiu, L., Wang, N., Ma, Y., Liu, Y., & Wen, D. (2018). Saponins from Boussingaultia gracilis prevent obesity and related metabolic impairments in diet-induced obese mice.\u00a0Food & Function.
\nOtt, R., Melchior, K., Stupin, J. H., Ziska, T., Schellong, K., Henrich, W., … & Plagemann, A. (2018). Reduced insulin receptor expression and altered DNA methylation in fat tissues and blood of women with GDM and offspring.\u00a0The Journal of Clinical Endocrinology & Metabolism.
\nYue, S. J., Liu, J., Wang, A. T., Meng, X. T., Yang, Z. R., Peng, C., … & Yan, D. (2018). Berberine alleviates insulin resistance by reducing peripheral branched-chain amino acids.\u00a0American Journal of Physiology-Endocrinology and Metabolism.
\nShah, A. P., Johnson, M. D., Fu, X., Boersma, G. J., Shah, M., Wolfgang, M. J., … & Baraban, J. M. (2019). Deletion of translin (Tsn) induces robust adiposity and hepatic steatosis without impairing glucose tolerance.\u00a0International Journal of Obesity, 1.
\nLi, M., Wang, M., Liu, Y., Huang, S., Yi, X., Yin, C., … & Xiao, Y. (2019). TNF\u2010\u03b1 Upregulates IKK\u03b5 Expression via the Lin28B\/let\u20107a Pathway to Induce Catecholamine Resistance in Adipocytes.\u00a0Obesity.
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\nWang, N., Ma, Y., Liu, Z., Liu, L., Yang, K., Wei, Y., … & Wen, D. (2019). Hydroxytyrosol prevents PM2. 5-induced adiposity and insulin resistance by restraining oxidative stress related NF-\u03baB pathway and modulation of gut microbiota in a murine model.\u00a0Free Radical Biology and Medicine,\u00a0141, 393-407. Zani, S. (2019).
\nExploratory study of egg white hydrolysate mechanisms of action in insulin sensitive tissues of high fat diet-induced insulin resistant rats.\u00a0https:\/\/doi.org\/10.7939\/r3-vpcv-w240
\nVianello, E., Dozio, E., Bandera, F., Froldi, M., Micaglio, E., Lamont, J., … & Schmitz, G. (2020). Correlative study on impaired prostaglandin E2 regulation in EAT and maladaptive cardiac remodeling via EPAC2 and ST2 signaling in overweight CVD subjects.\u00a0International Journal of Molecular Sciences,\u00a021, 520.
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\nWang, Y., Jiang, H. F., Liu, B. B., Chen, L. L., Wang, Y., Liu, X. Y., … & Wu, X. F. (2021). Brown Adipose Tissue Activation Is Involved in Atherosclerosis of ApoE\u2212\/\u2212 Mice Induced by Chronic Intermittent Hypoxia.\u00a0Frontiers in Cardiovascular Medicine,\u00a08.
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\nde Campos Zani, S. C., Son, M., Bhullar, K. S., Chan, C. B., & Wu, J. (2022). IRW (Isoleucine\u2013Arginine\u2013Tryptophan) Improves Glucose Tolerance in High Fat Diet Fed C57BL\/6 Mice via Activation of Insulin Signaling and AMPK Pathways in Skeletal Muscle.\u00a0Biomedicines,\u00a010(6), 1235.
\nCao, S., Pan, Y., Tang, J., Terker, A. S., Arroyo Ornelas, J. P., Jin, G. N., … & Zhang, M. Z. (2022). EGFR-mediated activation of adipose tissue macrophages promotes obesity and insulin resistance.\u00a0Nature Communications,\u00a013(1), 1-16.
\nLi, X., Zhang, X., Shen, Z., Chen, Z., Wang, H., & Zhang, X. (2022). GnRH receptor mediates lipid storage in female adipocytes via AMPK pathway.\u00a0Int J Med Sci,\u00a019(9), 1442-1450.
\nLiu, Z., Chen, T., Zhang, S., Yang, T., Gong, Y., Deng, H. W., … & Chen, Y. (2022). Discovery and functional assessment of a novel adipocyte population driven by intracellular Wnt\/\u03b2-catenin signaling in mammals.\u00a0bioRxiv, 2021-01.<\/p>\n

\"invent_list\"<\/p>\n

 <\/p>\n

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