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| Comparison of Tributyl phosphine (TBP) and DL-Dithiothreitol (DTT) for Two-dimensional Gel Electrophoresis(2-DE) of Qinchuan Cattle (Bos taurus) Longissimus Muscle Tissue |
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Abstract Appropriate reducing agent is very crucial for complete dissolution of protein and better results in two-dimensional gel electrophoresis (2-DE). Tributyl phosphine (TBP) is considered to be a more effective reducing agent than DL-Dithiothreitol (DTT) in general. In this study, we detailly compared DTT with TBP in term of 2-DE map quality and protein spot distribution of total proteins and sarcoplasmic proteins from Qinchuan cattle (Bos taurus) longissimus muscle tissue for the first time. The result implied that the reducing agent of DTT produced a higher resolution 2-DE gels with a clearer background than TBP. For the total proteins, DTT extracted more proteins with molecular weight ＞20 kD and protein volume ＜1 000 than TBP. For the sarcoplasmic proteins DTT promoted extraction of proteins with molecular weight＞50 kD and protein volume＜5 000 compared with TBP. DTT was more appropriate to 2-DE of Qinchuan cattle longissimus muscle tissue than TBP and this result wasn't consistent with traditional view. Further more, we increased TBP concentration and gave up the use of thiourea and the result wasn't improved. Through centrifugation protein precipitations appeared at room temperature. The short half-life of TBP might result in the poor effect in 2-DE. This study offers technical support for sample preparation and the proteomics research of Qinchuan cattle muscle tissue.
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Received: 09 September 2014
Published: 01 March 2015
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