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A two-dimensional electrophoresis protocol suitable for <i>Medicago truncatula</i> leaf proteome


Weimin Li
Xin Zhang
Wen Dong
Rong Jin
Yuping Wang
Di Wang
Feng Zhang
Zhenwu Wei

Abstract

Medicago truncatula leaves were used as the experimental materials. Total proteins of leaves were extracted by trichloracetic acid (TCA)-acetone method and proteins had a better separation using gel strips, forming an immobilized non-linear 3 to 10 pH gradient focusing 123,000 vhr combined with 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The gels were stained with Coomassie Brilliant Blue G-250 and digitalized gels were analyzed using the PDquest 8.0.1 software. The results indicated that 931 protein dots were detected in the gel. A technology suitable for the M. truncatula leaves protein extraction by TCA/acetone and a protocol for two-dimensional electrophoresis (2-DE) was established, which provides technical support for M. truncatula leaf proteome research.

Keywords: Medicago truncatula, proteome, two-dimensional polyacrylamide gel electrophoresis (2-DE), isoelectrofocusing (IEF)

African Journal of Biotechnology Vol. 12(3), pp. 233-237

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eISSN: 1684-5315