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| Optimization and Problem Analysis of Two-dimensional Electrophoresis Separation on Cashmere Goat(Capra cashmere) Skin Protein |
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Abstract To establish two-dimensional gel electrophoresis methods(2-DE) for cashmere goat skin, the experimental conditions of 2-DE were optimized, and various extraction and purification methods, different loading quantities and pH IPG strips, different isoelectric focusing were studied. The results showed that the process of grinding in liquid nitrogen and following by ultrasonication, the effect of sample purification with 2-D clean up kit, pH 4~7 17 cm pH IPG strip, the 450 μg loading quantity, isoelectric focusing process repeated several times in addition to salt step, maps were of good quality. Meanwhile, the common problems occurring in 2-DE experience were analyzed. The results indicated that the 2-DE maps of cashmere goat skin were obtained with high resolution and reproducibility, and can be used for the further proteome analysis. This study got a good 2-DE maps for cashmere goat skin protein, proved that it was feasible to conduct proteomics research for cashmere goat, and provided important technical information for researching development cycles and revealing origin and development law of skin hair follicle.
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Received: 23 August 2013
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