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77-86-1 Buffer TRIS,
Protein Extraction Buffer TRIS
Application of biological buffer TRIS (77-86-1) in plant protein extraction CAS.77-86-1 color white
Keywords: Biological buffer, TRIS, Tris, CAS77-86-1
The Chinese product name of Tris is Tris, CAS77-86-1. It is a white crystal or powder. Soluble in ethanol and water, slightly soluble in ethyl acetate and benzene, insoluble in ether and carbon tetrachloride, corrosive to copper and aluminum, and irritating chemicals.
Proteomics is to further understand the mechanism of life activities and the molecular mechanism of disease occurrence from the level of proteomics. In the research of proteomics, protein extraction is one of the most basic and critical links. Tris is the extraction of plant protein. Commonly used reagents, methods include Tris-HCl method, phenol method, Tris-acetone-phenol method. This article will analyze and summarize these methods.
1. Tris-HCl method
The effect of the protein spectrum extracted by the Tris-HCl method is better than that by the TCA acetone method. The protein extracted by the TCA acetone method is unevenly distributed in the small molecular weight region, the protein spots are not clear, and the horizontal and vertical stripes are more serious. The Tris-HCl method overcomes the above shortcomings and separates the acidity that is not easy to separate by the TCA acetone method. For protein, in addition to separating more medium molecular weight proteins, a lot of high molecular weight and low molecular weight proteins can also be obtained.
The Tris-HCl method is simple to operate, short in time and moderate in cost, with simple extraction steps, less protein loss, and good reproducibility of experimental results.
2. Phenol method
The phenol method takes advantage of the characteristics of Tris-saturated phenol: Phenol is a good solvent for proteins. During the sample preparation process, proteins and lipids are dissolved in the phenol phase, and soluble substances such as salts, nucleic acids, polyphenols and polysaccharides enter the water phase; Centrifugal force and centrifugal time can separate the dense sugar to the upper layer of the supernatant as much as possible; the protein in the phenol phase is precipitated by the methanol solution of ammonium acetate, and then washed with cold acetone several times to remove pigments and ammonium ions. Impurities.
This method is very effective in processing plant samples containing a large amount of interfering substances, but this method is relatively complicated and time-consuming.
The TCA acetone method is combined with the phenol method for protein extraction, which combines the advantages of the TCA acetone precipitation method and the phenol method. First wash the remaining TCA with a methanol solution containing ammonium acetate and raise the pH of the solution above 7.0, and then use a mixture of phenol and SDS to dissolve the protein. The combined extraction of phenol and SDS is more effective than the extraction of phenol and SDS buffer alone. The Tris-acetone-phenol method has achieved very good results in the protein extraction of many plants, and the background and bands of the electrophoretic pattern are very clear, and there are many protein spots. It can be widely used in the extraction of protein from plant leaves and fruits.
The extraction method is very time-saving, and the total extraction time is about 1 hour, which greatly shortens the time required for the traditional method to extract protein.
For the extraction of plant protein, not only the samples are diverse, but the ingredients contained in each plant are also different, and there are many interference factors, so there is no method that can be applied to various samples. In the experiment, we must first clarify the purpose, whether to obtain a target protein or all the proteins, and then select the extraction method according to the characteristics of the sample, and optimize it.
The above describes the application of TRIS in the extraction of plant protein, in addition to TRIS (Trishydroxymethylaminomethane) has many uses. It can be used as an intermediate for fosfomycin, as a vulcanization accelerator, cosmetics (cream, lotion), mineral oil, and paraffin emulsifier. It can also be used as an acid gas absorbent to prepare buffers, surfactants, emulsifiers and accelerators. Desheng has 15 years of experience in R&D and sales of TRIS, has a mature R&D team, has in-depth research on TRIS, and can solve technical problems for customers. If you need it, you can call for consultation.
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