Establishment of Two-dimensional Gel Electrophoresis (2-DE) for Coelomic Fluid in Apostichopus japonicus

doi:10.11924/j.issn.1000-6850.2013-3218

Abstract

Abstract: In order to use 2-DE analysis in the coelomic fluid proteins of Apostichopus japonicus. The protein extraction methods, sample volume, pH of IPG gel strips and isoelectric focusing programs were optimized to obtain best results. The results showed that the protein yield and purity were increased by improved TCA-acetone precipitation treatment. The resolution and reproducibility of 2-DE profiles were significantly improved by electing 8.5 cm ( pH 4-6.5) IPG gel strips, loading the sample 45 μg and seting 250 v, 7 mA isoelectric focusing for 30 min then 1000 v for 3 h. The result offered an important reference in screening differential expression protein and proteomics in Apostichopus japonicus.

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