A buffer for co-immunoprecipitation and its application

Abstract

The present invention discloses a buffer for co-immunoprecipitation and its application. The buffer for co-immunoprecipitation includes Tris-HCl, NaCl, NP-40 and PEG, wherein the Tris-HCl and NaCl are in The molar concentration in the buffer solution is 10-50 mM and 100-150 mM respectively. The present invention adopts mild NP-40 detergent and organic polymer PEG to affect the water environment around the protein and the interaction between proteins, promote the aggregation of macromolecules, help the interacting proteins to approach and combine, and make the proteins The interaction between them becomes specific, reducing non-specific reactions, so that the bands in the photos obtained by the co-immunoprecipitation test are clear, single and reliable, which helps to make a correct judgment on the test results and improve the quality of the co-immunoprecipitation test. The accuracy of the test results.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the field of molecular biology technology, in particular to a buffer for co-immunoprecipitation and its application. Background technique [0002] When people study the function of proteins, it is often necessary to lyse or break the tissue or cells to be tested so that the proteins in them can be released, so that one or more proteins can be obtained to study their functions and properties. One of the common methods used when lysing tissue or cells is to use a lysis buffer. Different formulas of lysates will affect the effect of protein acquisition, including whether the maximum amount of protein can be harvested, especially whether a relatively pure protein can be obtained. Therefore, it is very important to choose an appropriate cell lysate. [0003] After the total cell protein is obtained by cell lysis, it is necessary to obtain a certain target protein by immunoprecipitation ...

AI Technical Summary

Problems solved by technology

The above-mentioned cell lysates are commonly used in laboratories. Among them, SDS and Triton X-100 reagents have strong ability to lyse cells and dissolve proteins, but they are easy to denature proteins. Generally, they are not suitable for immunoprecipitation and co-immunoprecipitation reactions. It is suitable for obtaining total cell protein and used in experiments such as Western blot; the detergent NP-40 in RIPA lysate is relatively mild, and the total cell protein obtained by lysing cells with it can be used for immunoprecipitation and co-immunoprecipitation reactions, but it is not suitable for certain The purification of some proteins and the study of the interaction of some proteins are better, but the effect is poor for other proteins, with more non-specific reactions, more bands shown in the photos, and even a large blurred area, which makes researchers unable to Judging the experimental results

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