| Message: | Tris-HCl method
The effect of Tris-HCl method was better than TCA acetone method. The results showed that the protein extracted by TCA acetone method was unevenly distributed in the low molecular weight area, the protein spots were not clear, and the horizontal and vertical stripes were more serious. Tris-HCl method overcame the above shortcomings, and separated the acidic protein which was difficult to be separated by TCA acetone method. In addition to separating more middle molecular weight protein, it also obtained a lot of high molecular weight and low molecular weight protein.
Phenol method
The phenol method takes advantage of the characteristics of Tris saturated phenol: phenol is a good solvent for protein; in the process of sample preparation, protein and lipid are dissolved in phenol phase, and soluble substances such as salt, nucleic acid, polyphenol and polysaccharide enter into water phase; by increasing centrifugal force and centrifugal time, the dense sugar can be separated from the upper layer of supernatant as much as possible; the protein in phenol phase is dissolved in methanol of ammonium acetate The pigment and ammonium ion can be removed by precipitating the solution and cleaning the protein with cold acetone for many times. |
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