A Universal Polyacrylamide Gel Protein Rapid Separation and Staining Kit

Abstract

The invention belongs to the field of separating and dyeing of protein molecules and particularly relates to a general type kit for fast separating and dyeing polyacrylamide gel protein. The kit contains seven reagents to implement the separating and dyeing of the protein. The kit is simple in operation, the preparation process of the reagents is simple, the steps of separating and dyeing the protein are simplified, the prepared SDS-PAGE gel is uniform in coagulation, unapparent in electrophoresisfringe effect and remarkable in dyeing effect, and therefore, the general type kit for fast separating and dyeing the polyacrylamide gel protein is applicable to the follow-on experiments of the researchers.

Description

technical field [0001] The invention belongs to the field of separation and staining of protein molecules, in particular to a general-purpose polyacrylamide gel rapid protein separation and staining kit. Background technique [0002] With the development of proteomics, protein separation and identification research is an important research content in the field of biological sciences. There are many methods for protein separation at present, and the method of separating proteins by using SDS-PAGE discontinuous buffer system is widely used in the fields of life science, medicine and health. However, there are still many problems to be solved during the use of SDS-PAGE: [0003] ① The repeatability of the electrophoresis results is poor: due to the difficulty in mixing the colloid, the coagulation time of the polymerization is long, and the coagulation is uneven, forming a "smiling" belt with upturned sides and a concave center; [0004] ② Poor quality uniformity of electroph...

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Problems solved by technology

[0003]①The repeatability of the electrophoresis results is poor: due to the difficulty of colloidal mixing, the polymerization coagulation time is longer, and the coagulation is uneven, forming a "smile" with upturned sides and a concave middle "Type belt;

[0004]② Poor quality uniformity of the electrophoresis carrier SDS-PAGE gel: 1) The stacking gel is not concentrated well, the reason is that the dose of TEMED and APS is insufficient or invalid; 2) The stacking gel is concentrated The speed is faster. The reason is that the amount of APS and TEMED is too much. At this time, the colloid will be too hard and easy to crack, and it is easy to burn the gel during electrophoresis;

[0005]③The glue making time is long

However, the preparation method of Coomassie Brilliant Blue dye is cumbersome, long preparation time, short storage period (1 month failure), large dosage, long dyeing time, deep dyeing background and other problems, which not only increase the workload of researchers and waste of reagents, Moreover, it pollutes the environment and further increases carbon emissions.

[0008] In view of the above problems, although there have been many countermeasures, such as the invention of "Biomacromolecule Concentration and Desalination Method and Kit (CN102072836A)" for gel preparation, etc., it has solved polyacrylamide method concentration or desalination, gel recovery, However, problems such as uneven coagulation of polyacrylamide, ready-to-use reagents of TEMED and APS, and long gel-making time are still unsolved; and inventions for protein staining such as "a Coomassie brilliant blue staining method and its Special gel fixative solution (CN1904578A)" and "a Coomassie Brilliant Blue G250 staining method and its special solution (CN102288470A)" solved the problems of Coomassie Brilliant Blue staining method with deep staining background, cumbersome operation and improved sensitivity, but Coomassie Brilliant Blue Problems such as complex configuration of dye solution, short storage period, and long dyeing time have not been resolved

Moreover, from the currently published inventions, the protein separation and staining analysis processes are independent, and there is no kit that can integrate SDS-PAGE protein separation and staining tests

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