Polyacrylamide gel for separating small-molecular-weight proteins and preparation method of polyacrylamide gel

A polyacrylamide gel and protein separation technology, which is applied in the preparation methods of peptides, chemical instruments and methods, peptides, etc., can solve the problem of high cost of buffer reagents, and achieve the effects of reduced preparation costs, good separation, and reduced use costs.

Active Publication Date: 2019-12-03
武汉赛维尔生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to solve the shortcoming of the high cost of buffer reagents for the separation of small molecular weight proteins in the prior art, and propose a polyacrylamide gel for small molecular weight protein separation and its preparation method

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  • Polyacrylamide gel for separating small-molecular-weight proteins and preparation method of polyacrylamide gel
  • Polyacrylamide gel for separating small-molecular-weight proteins and preparation method of polyacrylamide gel

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Embodiment 1

[0032] A polyacrylamide gel for the separation of small molecular weight proteins, the gel includes acrylamide, N,N-methylenebisacrylamide, ultrapure water, gel buffer, glycerin, ammonium persulfate solution and tetramethyl ethylenediamine, the gel buffer includes boric acid and Tris-HCl solution, the pH of the gel buffer is 7.0, the concentration of Tris is 50-200mM, and the concentration of boric acid is 50-200mM.

[0033] The present invention also provides a method for preparing polyacrylamide gel used for the separation of small molecular weight proteins, comprising the following steps:

[0034] Step 1: prepare 30% acrylamide stock solution, wherein the mass ratio of acrylamide to N,N-methylene bisacrylamide is 29:1, the specific implementation steps are as follows:

[0035] A. First, weigh 29g of acrylamide and 1g of N,N-methylenebisacrylamide into the mixing container;

[0036] B. Then, add ultrapure water to the container, so that acrylamide and N,N-methylenebisacryla...

Embodiment 2

[0058] As another preferred embodiment of the present invention, the difference from the previous embodiment is that it is a polyacrylamide gel for the separation of small molecular weight proteins, which includes acrylamide, N, N-methylenebisacrylamide , ultrapure water, gel buffer, glycerol, ammonium persulfate solution and tetramethylethylenediamine, the gel buffer includes boric acid and Tris-HCl solution, the pH of the gel buffer is 7.0, and the concentration of Tris is 50-200mM, the concentration of boric acid is 50-200mM.

[0059] The present invention also provides a method for preparing polyacrylamide gel used for the separation of small molecular weight proteins, comprising the following steps:

[0060] Step 1: prepare 30% acrylamide stock solution, wherein the mass ratio of acrylamide to N,N-methylene bisacrylamide is 29:1, the specific implementation steps are as follows:

[0061] A. First, weigh 29g of acrylamide and 1g of N,N-methylenebisacrylamide into the mixi...

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Abstract

The invention belongs to the technical field of protein polyacrylamide gel electrophoresis, and especially relates to a polyacrylamide gel for small molecular weight protein separation. The polyacrylamide gel comprises acrylamide, N, N-methylene bisacrylamide, ultrapure water, a gel buffer solution, glycerin, an ammonium persulfate solution and tetramethylethylenediamine; the gel buffer solution comprises boric acid and a Tris-HCl solution, and the pH value of the gel buffer solution is 7.0. The polyacrylamide gel is prepared by taking Tris-boric acid as a buffer system; small molecular proteins can be well separated; protein strips are clear and sharp; the separation effect is equivalent to that of currently common small molecular protein separation gel Tris-Tricine polyacrylamide gel; the separation effect is even better than that of Tris-Tricine; the use cost of reagents is greatly reduced; and a better separation effect can be obtained.

Description

technical field [0001] The invention relates to the technical field of protein polyacrylamide gel electrophoresis, in particular to a polyacrylamide gel for separating small molecular weight proteins and a preparation method thereof. Background technique [0002] Polyacrylamide gel electrophoresis is a technology that uses polyacrylamide gel as the medium for protein separation analysis and detection. Polyacrylamide gel is a macromolecule polymerized by acrylamide and N,N-methylenebisacrylamide. , the network structure formed by the polymerization of amide side chains has the function of molecular sieve, and the pore size of the network formed by the gel is inversely proportional to the gel concentration. The higher the concentration, the smaller the pore size, and the stronger the ability to separate small molecular weight proteins; Smaller, the larger the pore size, the better the effect of separating macromolecular proteins. When a protein is electrophoresed in a polyacry...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08F220/56C08F222/38C08J3/075C08K5/17C08K5/053C08K3/38C07K1/26
CPCC08F220/56C08J3/075C08K5/17C08K5/053C08K3/38C07K1/26C08F222/385
Inventor 杨颂王绪德张高英
Owner 武汉赛维尔生物科技有限公司
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