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Usually buffalo cannot precisely express estrus signals. Hence, there is a need for concise methods to detect the time of estrus to ensure the success of artificial insemination. Therefore, we have established a reference proteome map on the whole saliva of buffalo during their estrous cycle with special reference to estrus. Nearly 12 bands have been observed using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS\u2010PAGE) of whole saliva. Collectively, 179 proteins are identified with respect to different phases of the estrous cycle using mass spectrometry. On the whole, 37 proteins are exclusively expressed in the estrus phase, which include \u03b2\u2010enolase, Toll\u2010like receptor (TLR) 4, clusterin, lactoperoxidase, serotransferrin, TGM3, UBA6, and transducin. Among the proteins, \u03b2\u2010enolase and TLR 4 were validated, and their specific expression was found during estrus as compared to other phases using immunoblot. The functional annotation reveals many as binding proteins in the estrus saliva when compared to the other phases. The present findings conclude that the proteomic approach adopted to identify the proteins from buffalo saliva around the estrous cycle may provide a new tool for screening the estrus phase. The results further conclude that the specific expression of \u03b2\u2010enolase and TLR 4 can be taken as the indicator of estrus in buffalo.\u2014Muthukumar, S., Rajkumar, R., Rajesh, D., Saibaba, G., Liao, C.\u2010C., Archunan, G., Padmanabhan, P., Gulyas, B., Exploration of salivary proteins in buffalo: an approach to find marker proteins for estrus. FASEB J. 28, 4700\u20134709 (2014).\n                    <jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"http:\/\/www.fasebj.org\">www.fasebj.org<\/jats:ext-link>\n                  <\/jats:p>","DOI":"10.1096\/fj.14-252288","type":"journal-article","created":{"date-parts":[[2014,8,12]],"date-time":"2014-08-12T02:56:50Z","timestamp":1407812210000},"page":"4700-4709","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["Exploration of salivary proteins in buffalo: an approach to find marker proteins for estrus"],"prefix":"10.1096","volume":"28","author":[{"given":"Subramanian","family":"Muthukumar","sequence":"first","affiliation":[{"name":"Center for Pheromone Technology Department of Animal Science Bharathidasan University Tiruchirappalli Tamil Nadu India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ramalingam","family":"Rajkumar","sequence":"additional","affiliation":[{"name":"Institute of Medical Biology A\u2010STAR Singapore"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Durairaj","family":"Rajesh","sequence":"additional","affiliation":[{"name":"Center for Pheromone Technology Department of Animal Science Bharathidasan University Tiruchirappalli Tamil Nadu India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ganesan","family":"Saibaba","sequence":"additional","affiliation":[{"name":"Center for Pheromone Technology Department of Animal Science Bharathidasan University Tiruchirappalli Tamil Nadu India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chen\u2010Chung","family":"Liao","sequence":"additional","affiliation":[{"name":"Proteomics Research Center National Yang\u2010Ming University Taipei Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Govindaraju","family":"Archunan","sequence":"additional","affiliation":[{"name":"Center for Pheromone Technology Department of Animal Science Bharathidasan University Tiruchirappalli Tamil Nadu India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Parasuraman","family":"Padmanabhan","sequence":"additional","affiliation":[{"name":"Lee Kong Chian School of Medicine Nanyang Technological University Singapore"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Balazs","family":"Gulyas","sequence":"additional","affiliation":[{"name":"Lee Kong Chian School of Medicine Nanyang Technological University Singapore"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2014,8,11]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1177\/10454411930040030101"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1749-6632.1993.tb18337.x"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.bdj.4807861"},{"key":"e_1_2_9_5_1","doi-asserted-by":"crossref","first-page":"196","DOI":"10.1177\/154405910408300301","article-title":"Dental, oral, and craniofacial research: the view from the NIDCR","volume":"83","author":"Tabak L. 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