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Buffer system for polyacrylamide precast gel for electrophoresis

A polyacrylamide and buffer system technology, which is applied in the direction of material analysis, instruments, and analysis materials by electromagnetic means, can solve the problems of low experimental efficiency, cumbersome procedures, easy to cause pollution, etc., so as to reduce protein modification and improve analysis. Results, the effect of fast electrophoresis

Inactive Publication Date: 2010-09-08
杭州吉来生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The purpose of the present invention is to overcome the defects of the above-mentioned prior art that the laboratory preparation process is cumbersome, time-consuming, easy to cause pollution, low experimental efficiency, and the shelf life of the existing prefabricated electrophoresis gel cannot meet long-term transportation, and provide a low-cost manufacturing method. Buffer system system for precast gels that are inexpensive, uniform and fast, and can be stored for more than 12 months to maintain protein separation ability and gel shape

Method used

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  • Buffer system for polyacrylamide precast gel for electrophoresis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] A buffer system for polyacrylamide prefabricated gel for electrophoresis, comprising: anodic electrophoresis tank buffer, cathodic electrophoresis tank buffer, separation gel and concentrated gel.

[0025] Concentrating gel and separating gel contain: 200mmol / L trishydroxymethyl aminomethane and 0.2% (w / v) sodium lauryl sulfate, titrate pH to 6.8 with citric acid (Citrate);

[0026] Contain 100mmol / L trihydroxymethylaminomethane-N-(2-hydroxyethyl)piperazine-N'-2-ethanesulfonic acid and 0.2% (w / v) Sodium lauryl sulfate, the pH value is adjusted to 7.8.

Embodiment 2

[0028] The composition of the buffer system for polyacrylamide precast gels for electrophoresis:

[0029] Gel storage solution: acrylamide (Acr) 28.38g, bisacrylamide (Bir) 1.62g.ddH 2 O, dilute to 100ml, filter and store at 4°C in the dark.

[0030] Shrink gel, 8% gel stock solution, 100mmol / L trishydroxymethyl aminomethane, 0.1% (w / v) sodium lauryl sulfate, and titrate pH to 6.5 with citric acid;

[0031] Separating gel, 4% gel stock solution, 100mmol / L trishydroxymethylaminomethane, 0.1% (w / v) sodium lauryl sulfate, and titrate pH to 6.5 with citric acid;

[0032] Anode electrophoresis tank buffer: 1mol / LH 3 PO 4 , 67ml H 3 PO 4 with ddH 2 O, dilute to 1L, add 80mmol / L Tris-HEPES and 0.1% (w / v) sodium lauryl sulfate;

[0033] Cathode electrophoresis tank buffer: 1mol / L NaOH, ddH for 40g NaOH 2 O, dilute to 1L, add 80mmol / L Tris-HEPES and 0.1% (w / v) sodium lauryl sulfate;

[0034] The pH was adjusted to 7.5.

Embodiment 3

[0036] Concentrating gel and separating gel contain: 300mmol / L trishydroxymethylaminomethane and 0.4% (w / v) sodium lauryl sulfate, and titrate the pH to 6.9 with citric acid;

[0037] Contain 120mmol / L trihydroxymethylaminomethane-N-(2-hydroxyethyl)piperazine-N'-2-ethanesulfonic acid and 0.4% (w / v) Sodium lauryl sulfate, the pH value is adjusted to 8.

[0038] The present invention can be applied to commercially available electrophoresis tanks or electrophoresis apparatuses, and more preferably to be used in disposable prefabricated ready-to-use overall electrophoresis devices.

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Abstract

The invention relates to a buffer system for polyacrylamide precast gel for electrophoresis, and belongs to the technical field of gel electrophoresis. The buffer system comprises anode electrophoresis tank buffer solution, cathode electrophoresis tank buffer solution, separating gel and concentrated gel and is characterized in that: the concentrated gel and the separating gel comprises 100 to 300mmol / L of trihydroxymethyl aminomethane and 1 to 4g / L of lauryl sodium sulfate and is titrated by using nitric acid to adjust the pH value to be between 6.5 and 6.9; and the anode electrophoresis tank buffer solution and the cathode electrophoresis tank buffer solution comprise 80 to 120mmol / L of trihydroxymethyl aminomethane-N-(2-hydroxyethyl) piperazine-N'-2-ethane sulfonic acid and 1 to 4g / L of lauryl sodium sulfate and the pH value is adjusted to be between 7.5 and 8. The system has the advantages of enhancing the stability of a gel matrix and the buffer solution, not reducing the performance of the gel matrix and the buffer solution when the gel matrix and the buffer solution are stored in a refrigerator for more than one year and improving the analysis result of Western blot.

Description

technical field [0001] The invention belongs to the technical field of gel electrophoresis, in particular to a polyacrylamide gel buffer system used for prefabricated gel electrophoresis. Background technique [0002] Polyacrylamide gel electrophoresis (PAGE) is an electrophoresis method using polyacrylamide gel as a supporting medium. On this support medium, the separated substance can be separated according to the molecular size and molecular charge. Since polyacrylamide gel is a kind of artificially synthesized substance, the concentration ratio of monomers can be adjusted before polymerization to form different degrees of cross-linked structures, and its porosity can be changed in a wide range, and substances can be separated according to the needs. The size of the molecule, choose the appropriate gel composition and concentration, so that it has a better resolution. Polyacrylamide gel electrophoresis is the most commonly used electrophoresis method for protein separat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/447B01D57/02
Inventor 汪弋
Owner 杭州吉来生物技术有限公司
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