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Application of Tris (77-86-1) in Plant Protein Extraction

2017-08-07 来源:亚科官网
7 August 2017
  Proteomics is to further understand the mechanism of life activities and the molecular mechanism of disease occurrence from the level of the proteome, and in the research of proteomics, protein extraction is one of the most basic and most critical aspects. Tris is a common agent in the experiment of plant protein extraction, the methods include Tris-HCl, Tris-phenol, Tris-acetone-phenol. These methods will be summarized in this article.
  The effect of Tris-HCl on the extraction of protein was better than that of TCA-acetone. TCA-acetone extraction of protein is uneven in the small molecular weight region, protein spots are not clear, horizontal stripes and vertical stripes are relatively serious, while Tris-HCl overcomes the above shortcomings and separates the acidic protein that is difficult to separate by TCA-acetone method. In addition to the separation of more moderate molecular weight protein, tris-HCl also could get a lot of high molecular weight and low molecular weight protein.
  Tris-HCl method has the advantages of short extraction time, moderate cost, simple extraction step, less protein loss, and reproducible experimental result.
  Tris-phenol use the characteristics of Tris-saturated phenol: phenol is a good solvent for protein, , protein and lipids are dissolved in phenolic in the preparation of the sample, while salt, nucleic acid, polyphenols and polysaccharides and other soluble substances are into the water phase; By increasing the centrifugal force and centrifugation time, it is possible to leave the density of sugar to the upper layer of the supernatant as much as possible; Protein in phenol phase could precipitate through the methanol solution of ammonium acetate, and then cleaning the protein with cold acetone several times, pigments and ammonium ions and other impurities can be removed.
  This method is very effective in handling plant samples containing a large amount of interfering substances, but it is relatively complex and time consuming.
  The combination of TCA-acetone and tris-phenol for the extraction of protein, has the advantages of TCA-acetone precipitation and tris-phenol. The residual TCA is first washed with a methanol solution containing ammonium acetate and adjust the pH of the solution to above 7.0, and a mixture of phenol and SDS is used to dissolve the protein. Phenol-SDS combined extraction is more effective than phenol, SDS buffer extraction alone. Tris-acetone-phenol method has achieved very good results in protein extraction of many plants, and the background and strip in electrophoretic pattern are very clear, it can be widely used in plant leaves and fruit protein extraction.
  The extraction method is very time-saving, the total extraction time is about 1h, greatly less than that of the traditional method.
  For the extraction of plant protein, the sample is diverse, and each plant contains different components, and there are many interfere factors, so no one method can be applied to a variety of samples. In the experiment, the purpose (to obtain a target protein or all the protein) must be cleared firstly, and then select the extraction method according to the characteristics of the sample.
Related links: Tris
Extraction of Total Proteins in Cells and Tissues

Edited by Suzhou Yacoo Science Co., Ltd.