CN-121972040-A - Simple and rapid multi-concentration glue preparation kit and glue preparation method

Abstract

The invention relates to the technical field of biochemical experiments, and particularly discloses a simple and rapid multi-concentration glue preparation kit and a glue preparation method, wherein the glue preparation kit comprises lower glue, upper glue and a coagulant, the upper glue comprises upper glue A liquid and upper glue B liquid, the lower glue A liquid comprises lower glue A1 liquid, lower glue A2 liquid and lower glue B liquid, the lower glue A1 liquid, the lower glue A2 liquid and ultrapure water are mixed to form lower glue A liquid with different concentrations, a user only needs to refer to a simple proportioning table, the lower glue A1 liquid, the lower glue A2 liquid and the ultrapure water are mixed in different volumes, so that lower separation glue working liquids with various concentrations can be rapidly and accurately prepared on the basis of a general formula, the lower glue B liquid, the upper glue A liquid and the upper glue B liquid are kept fixed and universal, and variable independent adjustment and constant and uniform supply are ingeniously realized.

Inventors

• ZHOU FATING
• LI CHAOFU
• ZHU XIAODAN
• Que Wanshu

Assignees

• 重庆市急救医疗中心(重庆市第四人民医院、重庆市急救医学研究所)

Dates

Publication Date

20260505

Application Date

20251225

Claims (10)

1. 1. The simple and rapid multi-concentration glue preparation kit comprises lower glue, upper glue and a coagulant, and is characterized in that the upper glue comprises upper glue A liquid and upper glue B liquid; the lower layer glue A liquid comprises lower layer glue A1 liquid, lower layer glue A2 liquid and lower layer glue B liquid, wherein the lower layer glue A1 liquid, the lower layer glue A2 liquid and the ultrapure water are mixed to form lower layer glue A liquid with different concentrations.
2. 2. The simple and rapid multi-concentration glue preparation kit according to claim 1, wherein the lower glue A1 solution is a mixed mother solution of Acr and Bis; the lower layer adhesive A2 liquid comprises TEMED and ultrapure water; the lower layer glue B liquid comprises Tris-HCl and SDS; the upper layer adhesive A liquid comprises Acr-Bis, TEMED and ultrapure water; the upper layer glue B liquid comprises Tris-HCL, SDS and ultrapure water; the coagulant is ammonium persulfate.
3. 3. The simple and rapid multi-concentration glue preparation kit according to claim 2, wherein the concentration of the mixed mother liquor in the lower glue A1 solution is 30%, and the ratio of Acr to Bis is 29:1; in the lower layer adhesive A2 liquid, the ratio of TEMED to ultrapure water is 0.16:12; in the lower layer glue B solution, the pH value of Tris-HCl is 8.8, and SDS is 10%; In the upper layer adhesive A liquid, the concentration of Acr-Bis is 30%, and the ratio of Acr-Bis, TEMED and ultrapure water is 34:0.2:66; In the upper layer gum B liquid, the pH value of Tris-HCl is 6.8, and the ratio of Tris-HCL, SDS and ultrapure water is 26:2:72; in the coagulant, the concentration of ammonium persulfate is 10%.
4. 4. The simple and rapid multi-concentration glue preparation kit according to claim 3, wherein the lower glue A liquid is prepared by mixing lower glue A1 liquid, lower glue A2 liquid and ultrapure water according to the following volume ratio of 26:3.04:21.
5. 5. The simple and rapid multi-concentration glue preparation kit according to claim 3, wherein the lower glue A liquid is prepared by mixing lower glue A1 liquid, lower glue A2 liquid and ultrapure water according to the following volume ratio of 34:3.04:13.
6. 6. The simple and rapid multi-concentration glue preparation kit according to claim 3, wherein the lower glue A liquid is prepared by mixing lower glue A1 liquid, lower glue A2 liquid and ultrapure water according to the following volume ratio of 40:3.04:7.
7. 7. The simple and rapid multi-concentration glue preparation kit according to claim 3, wherein the lower glue A liquid is prepared by mixing lower glue A1 liquid, lower glue A2 liquid and ultrapure water according to the following volume ratio of 46:3.04:1.
8. 8. A simple and rapid multi-concentration glue preparation method, which is characterized by comprising the step of preparing separation glue with different concentrations by using the glue preparation kit of any one of claims 1-7.
9. 9. The method for preparing the simple and rapid multi-concentration adhesive according to claim 8, wherein the step of preparing the separation adhesive with different concentrations comprises the following steps: Step S1, preparing a coagulant; step S2, sequentially measuring lower layer adhesive A1 liquid, lower layer adhesive A2 liquid and ultrapure water, and preparing lower layer adhesive A liquid with corresponding concentration; step S3, taking lower layer glue A liquid, lower layer glue B liquid and coagulant, mixing to obtain lower layer glue, and mixing upper layer glue A liquid, upper layer glue B liquid and coagulant to obtain upper layer glue; S4, in the step S4, the mixed solution is quickly poured into a gel mold, and a layer of isopropanol is capped; and S5, standing at room temperature for 8-12 minutes, wherein a clear interface is observed to indicate that the polymerization is completed.
10. 10. The method for preparing the simple and rapid multi-concentration glue according to claim 9, wherein the concentration of the lower glue A is 8%, 10%, 12% or 14%.

Description

Simple and rapid multi-concentration glue preparation kit and glue preparation method Technical Field The application relates to the technical field of biochemical experiments, and particularly discloses a simple and rapid multi-concentration glue preparation kit and a glue preparation method. Background Polyacrylamide gel electrophoresis (SDS-PAGE) is a fundamental and critical technique in the fields of biochemistry and molecular biology for separating and identifying proteins. The technology is based on the fact that proteins are charged in the presence of SDS (sodium dodecyl sulfate) in proportion to their molecular weight, and separation by molecular weight is achieved by migration in a polyacrylamide gel matrix. The concentration of the gel (i.e., the total content of acrylamide monomers) is a core parameter for determining the separation range, namely, the low concentration gel (such as 8%) has larger pore size, which is beneficial to the separation of high molecular weight proteins, and the high concentration gel (such as 14%) has smaller pore size, which is more suitable for the effective resolution of low molecular weight proteins. Therefore, in practical research work, gels with different concentrations are often selected flexibly according to the molecular weight range of the target protein. At present, the preparation of SDS-PAGE gels in the laboratory mainly depends on the following methods: One is the traditional manual preparation method, which requires an experimenter to independently weigh various reagents such as acrylamide, methylene bisacrylamide, tris-HCI, SDS and the like, and accurately adjust the pH value. The method has the advantages of complicated steps, time consumption of 30-50 minutes, higher technical requirements on operators, large batch-to-batch difference easily caused by inaccurate weighing or pH adjustment deviation, and poor experimental repeatability. In addition, acrylamide monomers are neurotoxic and frequent weighing increases the health risk to the experimenter. The other is a commercial one-step method kit, namely products which simplify the glue preparation process and are proposed by companies such as elegance and the like are available on the market, and the products usually mix or split various components in an acrylamide/methylene bisacrylamide mixed solution, a buffer solution, an initiator (such as ammonium persulfate) and an accelerator (such as TEMED) in advance, so that the workload of weighing and preparing various independent solutions by experimenters is reduced, and the convenience and the speed of operation are improved to a certain extent. However, these products typically can only be formulated as a gel at a fixed concentration in a single kit (e.g., only 12% of the separation gel can be formulated). When one experimental study requires simultaneous analysis of multiple proteins with widely varying molecular weight ranges, or different experimental projects require gels of different separation ranges, researchers have to purchase and store multiple independent kits corresponding to different concentration specifications. The reagent purchasing cost is greatly increased, and the storage space of a laboratory refrigerator or freezer is occupied because of the need of storing the reagent boxes with various packaging specifications, so that the waste of resources and space is caused. Therefore, the existing glue preparation kit can rapidly and flexibly prepare the separation glue with different concentrations, so that the test operation time, the reagent cost and the reagent storage space are increased. Therefore, the present invention provides a simple and rapid multi-concentration glue dispensing kit and a glue dispensing method, so as to solve the above problems. Disclosure of Invention The invention aims to solve the problems of increased test operation time, increased reagent cost and wasted reagent storage space caused by the fact that the traditional reagent kit for preparing the adhesive rapidly and flexibly prepares the separating adhesive with different concentrations. In order to achieve the purpose, the basic scheme of the invention provides a simple and rapid multi-concentration glue preparation kit, which comprises lower glue, upper glue and a coagulant, wherein the upper glue comprises upper glue A liquid and upper glue B liquid; the lower layer glue A liquid comprises lower layer glue A1 liquid, lower layer glue A2 liquid and lower layer glue B liquid, wherein the lower layer glue A1 liquid, the lower layer glue A2 liquid and the ultrapure water are mixed to form lower layer glue A liquid with different concentrations. Further, the lower layer adhesive A1 liquid is mixed mother liquid of Acr and Bis; the lower layer adhesive A2 liquid comprises TEMED and ultrapure water; the lower layer glue B liquid comprises Tris-HCl and SDS; the upper layer adhesive A liquid comprises Acr-Bis, TEMED and ultrapure water; the upper layer glue B liqui