Preparation method of protein imprinting membrane regeneration solution

Abstract

The invention relates to the technical field of molecular imprinting, in particular to a preparation method of a protein imprinting membrane regeneration solution, which comprises the following steps: treating a protease K solution with guanidine hydrochloride to obtain a treated protease K solution; low-activity protease K and a composite salt solution are mixed and prepared, the pH value is adjusted, a western blot membrane regeneration solution is obtained, the protease K can hydrolyze protein into small fragments of polypeptide, and therefore template protein can be eluted out of holes more easily, and a good elution effect is achieved; the protease K solution is treated by guanidine hydrochloride, so that the protease K can be defolded to a certain extent, the activity of the protease K and the conformation have relatively strong correlation, and the activity of the protease K can be reduced by inducing the protease K to perform the defolding treatment, so that more effective recognition sites are remained after elution; therefore, other proteins can be directly detected, and the method has better effect and elution efficiency.

Description

technical field [0001] The invention relates to the technical field of molecular imprinting, in particular to a method for preparing protein imprinting membrane regeneration solution. Background technique [0002] Western blotting is a commonly used test method in molecular biology, biochemistry and immunogenetics. Its basic principle is to color the cells or biological tissue samples treated by gel electrophoresis through specific antibodies, and analyze the coloring position and Gain in-depth information on how specific proteins are expressed in the cells or tissues being analyzed. [0003] The specific test method is to transfer the protein sample separated by polyacrylamide gel electrophoresis to a solid phase carrier. The solid phase carrier absorbs the protein in the form of non-covalent bonds, and can maintain the type of polypeptide separated by electrophoresis and its biological activity. Mutation, using the protein or polypeptide on the solid phase carrier as the ...

AI Technical Summary

Problems solved by technology

[0004] After completing the combination of primary antibody and secondary antibody and subsequent chemiluminescent detection in western blotting, sometimes other proteins need to be detected for comparison. The special components in the Western blot membrane regeneration solution can dissociate the combination of antigen and antibody on the membrane, but Affect the protein transferred to the membrane, and then use different antibodies for the next round of Western blot experiments to detect other proteins. When multiple protein detections are required, the traditional elution strength is too large, which often leads to solid phase support. The protein signal is weakened, and it contains mercaptoethanol, a highly irritating chemical substance. Careless operation will cause harm to the staff, and the operation time will be longer

[0005] For example, in the invention patent of publication number CN101833008 A, glycine and sodium dodecylsulfonate are used to prepare protein imprinted membrane regeneration solution, which has a good elution effect, but the elution strength is too large, which often leads to protein on the solid phase carrier. The signal is weakened and the antigen is easily destroyed

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