Buffer Optimization for Bovine Longissimus Muscle Tissues: Proteome Analysis of Korean Native Cattle Using 2-Dimensional Gel Electrophoresis

• Published in: Food science and biotechnology Vol. 19; no. 4; pp. 1107 - 1112
• Main Authors: Lee, Hong-Gu, Han, Jeung-A, Lee, Ki-Bum, Kim, Eun-Bea, Jin, Yong-Cheng, Oh, Jin-Ju, Hwang, Jin-Hee, Kang, Han-Suk, Kim, Sang-Hun, Seo, Kang-Suk, Kang, Sang-Kee, Choi, Yun-Jaie
• Format: Journal Article
• Language: Korean
• Published: 2010
• Subjects: 2-dimensional gel electrophoresis (2-DE); solubilization; 2-DE analysis; bovine longissimus muscle; resolution
• ISSN: 1226-7708; 2092-6456

Because beef contains a high proportion of marbling, we sought to optimize the solubilization of the longissimus muscle proteome using modified lysis and isoelectric focusing (IEF) rehydration buffers. Of the 3 lysis buffers tested, those containing a thiourea-urea mixture provided superior resolution, whereas that which contained only urea yielded consistently poor results. In addition, we found that the IEF rehydration buffer containing a thiourea-urea mixture, a high dithiothreitol level, and Pharmalyte was able to generate sharper 2-dimensional (2-D) gels, whereas that which contained immobilized pH gradient (IPG) buffer had no an effect on protein resolution. The number of different protein spots between the muscle-development stage (11 months) and the fat-development stage (17 months) in Hanwoo steers was 39, whereas the use of normal buffer resulted in a detection of a difference of only 7 protein spots. This study provides a methodological tool for studying the proteome of bovine longissimus muscle with a high fat content using 2-D gel electrophoresis (2-DE) gels.