Comparison of protein sample preparation methods of two-dimensional electrophoresis for Skeletonema costatum

WANG Xiu-xiu; CHEN Ji-xin; HUANG Bang-qin

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Journal of Marine Sciences ›› 2012, Vol. 30 ›› Issue (4) : 72-77.

Comparison of protein sample preparation methods of two-dimensional electrophoresis for Skeletonema costatum

WANG Xiu-xiu, CHEN Ji-xin, HUANG Bang-qin^*

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Abstract

Lysis buffer method and trichloroacetic acid ( TCA)-acetone precipitation method both are the common methods in algal protein abstraction. We compared the 2-DE maps of these two protein extraction methods to find out the best protocol for Skeletonema costatum. As a result,the Lysis buffer-TCA-acetone method had better effect than the other two lysis buffer methods (lysis buffer-microcon &. lysis buffer-2-D clean-up kit) in removing the intracellular interferential factors, such as salt, nucleic acid, polyphenol and polysaccharide. The lysis buffer-TCA-acetone method and straight TCA-acetone precipitation method both represented clean background and clear protein dots in 2-DE maps while the later method showed better isoelect rofocusing results. We optimized the straight TCA-acetone precipitation methods. The TCA-acetone precipitation with 12 hours precipitation and a following ultrasonic cleaning process method was confirmed to be the appropiate sample preparation method for S. costatum for two-dimensional electrophoresis.

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Skeletonema costatum / proteomics / two-dimensional electrophoresis / protein extraction method

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WANG Xiu-xiu, CHEN Ji-xin, HUANG Bang-qin. Comparison of protein sample preparation methods of two-dimensional electrophoresis for Skeletonema costatum[J]. Journal of Marine Sciences. 2012, 30(4): 72-77

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