{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,16]],"date-time":"2025-10-16T00:58:26Z","timestamp":1760576306440,"version":"build-2065373602"},"reference-count":31,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2017,11,20]],"date-time":"2017-11-20T00:00:00Z","timestamp":1511136000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"JSPS","award":["JP24700407"],"award-info":[{"award-number":["JP24700407"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Electrophoresis"],"published-print":{"date-parts":[[2018,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Phosphorylation analysis by using phos\u2010tag technique has been reported to be suitable for highly sensitive quantification of smooth muscle myosin regulatory light chain (LC<jats:sub>20<\/jats:sub>) phosphorylation. However, there is another factor that will affect the sensitivity of phosphorylation analysis, that is, protein extraction. Here, we optimized the conditions for total protein extraction out of trichloroacetic acid (TCA)\u2010fixed tissues. Standard SDS sample buffer extracted less LC<jats:sub>20<\/jats:sub>, actin and myosin phosphatase targeting subunit 1 (MYPT1) from TCA\/acetone treated ciliary muscle strips. On the other hand, sample buffer containing urea and thiourea in addition to lithium dodecyl sulfate (LDS) or SDS extracted those proteins more efficiently, and thus increased the detection sensitivity up to 4\u20135\u00a0fold. Phos\u2010tag SDS\u2010PAGE separated dephosphorylated and phosphorylated LC<jats:sub>20<\/jats:sub>s extracted in LDS\/urea\/thiourea sample buffer to the same extent as those in standard SDS buffer. We have concluded that LDS (or SDS) \/urea\/thiourea sample buffer is suitable for highly sensitive phosphorylation analysis in smooth muscle, especially when it is treated with TCA\/acetone.<\/jats:p>","DOI":"10.1002\/elps.201700394","type":"journal-article","created":{"date-parts":[[2017,10,26]],"date-time":"2017-10-26T09:36:52Z","timestamp":1509010612000},"page":"326-333","source":"Crossref","is-referenced-by-count":9,"title":["Addition of urea and thiourea to electrophoresis sample buffer improves efficiency of protein extraction from TCA\/acetone\u2010treated smooth muscle tissues for phos\u2010tag SDS\u2010PAGE"],"prefix":"10.1002","volume":"39","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4270-5829","authenticated-orcid":false,"given":"Kosuke","family":"Takeya","sequence":"first","affiliation":[{"name":"Department of Physiology Asahikawa Medical University  Hokkaido Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Toshiyuki","family":"Kaneko","sequence":"additional","affiliation":[{"name":"Department of Physiology Asahikawa Medical University  Hokkaido Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Motoi","family":"Miyazu","sequence":"additional","affiliation":[{"name":"Department of Physiology Asahikawa Medical University  Hokkaido Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Akira","family":"Takai","sequence":"additional","affiliation":[{"name":"Department of Physiology Asahikawa Medical University  Hokkaido Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2017,11,20]]},"reference":[{"doi-asserted-by":"publisher","key":"e_1_2_8_2_1","DOI":"10.1016\/S0968-0004(00)01712-6"},{"doi-asserted-by":"publisher","key":"e_1_2_8_3_1","DOI":"10.1083\/jcb.109.6.2887"},{"doi-asserted-by":"publisher","key":"e_1_2_8_4_1","DOI":"10.1111\/j.1469-7793.1999.0805u.x"},{"key":"e_1_2_8_5_1","first-page":"1558","volume":"39","author":"Kerrick W. G.","year":"1980","journal-title":"Fed. Proc."},{"doi-asserted-by":"publisher","key":"e_1_2_8_6_1","DOI":"10.1016\/S0021-9258(20)65117-1"},{"doi-asserted-by":"publisher","key":"e_1_2_8_7_1","DOI":"10.1016\/S0021-9258(18)43729-5"},{"doi-asserted-by":"publisher","key":"e_1_2_8_8_1","DOI":"10.1016\/S0021-9258(18)52353-X"},{"doi-asserted-by":"publisher","key":"e_1_2_8_9_1","DOI":"10.1074\/mcp.T500024-MCP200"},{"doi-asserted-by":"publisher","key":"e_1_2_8_10_1","DOI":"10.1152\/ajprenal.00060.2008"},{"doi-asserted-by":"publisher","key":"e_1_2_8_11_1","DOI":"10.1113\/jphysiol.2008.168252"},{"doi-asserted-by":"publisher","key":"e_1_2_8_12_1","DOI":"10.1113\/jphysiol.2010.187146"},{"doi-asserted-by":"publisher","key":"e_1_2_8_13_1","DOI":"10.1074\/jbc.M112.371609"},{"doi-asserted-by":"publisher","key":"e_1_2_8_14_1","DOI":"10.1038\/ki.2014.284"},{"doi-asserted-by":"publisher","key":"e_1_2_8_15_1","DOI":"10.1540\/jsmr.52.45"},{"key":"e_1_2_8_16_1","doi-asserted-by":"crossref","first-page":"1149","DOI":"10.1242\/jcs.112.8.1149","volume":"112","author":"Hayashi K.","year":"1999","journal-title":"J. Cell Sci."},{"doi-asserted-by":"publisher","key":"e_1_2_8_17_1","DOI":"10.1021\/pr025541x"},{"doi-asserted-by":"publisher","key":"e_1_2_8_18_1","DOI":"10.1002\/elps.200500757"},{"doi-asserted-by":"publisher","key":"e_1_2_8_19_1","DOI":"10.1371\/journal.pone.0124723"},{"doi-asserted-by":"publisher","key":"e_1_2_8_20_1","DOI":"10.1016\/S0021-9258(20)81332-5"},{"key":"e_1_2_8_21_1","first-page":"225","volume":"121","author":"Masato T.","year":"1997","journal-title":"J. Biochem."},{"doi-asserted-by":"publisher","key":"e_1_2_8_22_1","DOI":"10.1002\/pmic.201000472"},{"doi-asserted-by":"publisher","key":"e_1_2_8_23_1","DOI":"10.1023\/A:1026357009886"},{"doi-asserted-by":"publisher","key":"e_1_2_8_24_1","DOI":"10.1002\/pro.108"},{"doi-asserted-by":"publisher","key":"e_1_2_8_25_1","DOI":"10.1002\/elps.1150170503"},{"doi-asserted-by":"publisher","key":"e_1_2_8_26_1","DOI":"10.1002\/elps.1150180303"},{"doi-asserted-by":"publisher","key":"e_1_2_8_27_1","DOI":"10.1002\/1522-2683(200206)23:11<1745::AID-ELPS1745>3.0.CO;2-A"},{"doi-asserted-by":"publisher","key":"e_1_2_8_28_1","DOI":"10.1002\/elps.1150140110"},{"doi-asserted-by":"publisher","key":"e_1_2_8_29_1","DOI":"10.1007\/BF02609281"},{"doi-asserted-by":"publisher","key":"e_1_2_8_30_1","DOI":"10.1016\/0003-2697(86)90146-6"},{"doi-asserted-by":"publisher","key":"e_1_2_8_31_1","DOI":"10.1073\/pnas.76.1.111"},{"doi-asserted-by":"publisher","key":"e_1_2_8_32_1","DOI":"10.1002\/1522-2683(200105)22:9<1593::AID-ELPS1593>3.0.CO;2-6"}],"container-title":["ELECTROPHORESIS"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Felps.201700394","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/elps.201700394","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T18:16:26Z","timestamp":1760552186000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/elps.201700394"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,11,20]]},"references-count":31,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2018,1]]}},"alternative-id":["10.1002\/elps.201700394"],"URL":"https:\/\/doi.org\/10.1002\/elps.201700394","archive":["Portico"],"relation":{},"ISSN":["0173-0835","1522-2683"],"issn-type":[{"type":"print","value":"0173-0835"},{"type":"electronic","value":"1522-2683"}],"subject":[],"published":{"date-parts":[[2017,11,20]]}}}