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Protein electrophoresis staining solution, kit and staining method

A dyeing solution and protein technology, which is applied in biological testing, preparation of test samples, material analysis by electromagnetic means, etc., can solve the problems of long dyeing and decolorization, and achieve shortened dyeing time, low background dyeing, and dyeing speed. quick effect

Pending Publication Date: 2020-09-01
浙江玉安康瑞生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the problem of too long dyeing and decolorization time needs to be solved. The dyeing time of the current formula usually needs to reach more than 30 minutes to achieve better experimental results.

Method used

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  • Protein electrophoresis staining solution, kit and staining method
  • Protein electrophoresis staining solution, kit and staining method
  • Protein electrophoresis staining solution, kit and staining method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] 1. Preparation of Staining Solution

[0049] Weigh 250 mg Coomassie Brilliant Blue G-250 (CBB G-250), 300 mg Coomassie Brilliant Blue R-250 (CBB R-250), 100 ml glycerin, 1 ml essence, 20 g trehalose, 1 g Tween 20 , add deionized water to make up to 985ml, stir for 2-4 hours, after the Coomassie Brilliant Blue is fully dissolved, add 15ml of concentrated hydrochloric acid with a concentration of 38%, after mixing evenly, the final acidity of the solution is 14. Finally, store it in opaque bottles.

[0050] 2. SDS-PAGE electrophoresis band staining and decolorization

[0051] 1) After protein gel electrophoresis, wash the gel with 100ml of deionized water on a shaker in a plastic container for 1 min, discard the washed water, and repeat twice.

[0052] 2) Add 100ml of deionized water, heat it in a microwave oven to near boiling, but avoid boiling, then shake and wash on a shaker for 5 minutes. For gels with a thickness of 1.5mm, it is necessary to prolong the shaking w...

Embodiment 2

[0060] 1. Preparation of Staining Solution

[0061] Weigh 250mg of Coomassie Brilliant Blue G-250 (CBB G-250), 100ml of glycerin, 1ml of essence, 20g of trehalose, 1g of Tween 20, add deionized water to 985ml, stir for 2 to 4 hours, and wait for examination After the Mas Brilliant Blue is fully dissolved, add 15ml of concentrated hydrochloric acid with a concentration of 38%, and after mixing evenly, the final acidity of the solution is 14. Finally, store it in opaque bottles.

[0062] 2. SDS-PAGE electrophoresis band staining and decolorization

[0063] 1) After protein gel electrophoresis, wash the gel with 100ml of deionized water on a shaker in a plastic container for 1 min, discard the washed water, and repeat twice.

[0064] 2) Add 100ml of deionized water, heat it in a microwave oven to near boiling, but avoid boiling, then shake and wash on a shaker for 5 minutes. For gels with a thickness of 1.5mm, it is necessary to prolong the shaking washing time.

[0065] 3) S...

Embodiment 3

[0072] 1. Preparation of Staining Solution

[0073] Weigh 300 mg Coomassie Brilliant Blue R-250 (CBB R-250), 100 ml glycerin, 1 ml essence, 20 g trehalose, 1 g Tween 20, add deionized water to 985 ml, stir for 2-4 h, After the Coomassie Brilliant Blue is fully dissolved, add 15ml of concentrated hydrochloric acid with a concentration of 38%, and after mixing evenly, the final acidity of the solution is 14. Finally, store it in opaque bottles.

[0074] 2. SDS-PAGE electrophoresis band staining and decolorization

[0075] 1) After protein gel electrophoresis, wash the gel with 100ml of deionized water on a shaker in a plastic container for 1 min, discard the washed water, and repeat twice.

[0076] 2) Add 100 ml of deionized water, heat it in a microwave oven to near boiling, but avoid boiling, then shake and wash on a shaker for 5 minutes. For gels with a thickness of 1.5mm, it is necessary to prolong the shaking washing time.

[0077] 3) Shake and wash the gel with 100ml o...

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Abstract

The invention discloses a protein electrophoresis staining solution. The staining solution comprises Coomassie brilliant blue, concentrated hydrochloric acid and water. The invention further disclosesa method for dyeing a protein SDS polyacrylamide gel electrophoresis strip by using the dyeing solution. According to the invention, the formula of the protein electrophoresis staining solution is improved; the dyeing effect of the protein electrophoresis strip is improved; the dyeing and decolorizing speeds are increased, and meanwhile, the safety of the dyeing liquid to human bodies is ensured;and the staining solution and the method are particularly suitable for pollution-free, rapid and high-sensitivity dyeing of protein gels such as SDS-PAGE or non-denatured PAGE or detection of residual protein on PAGE gel after Western membrane transfer.

Description

technical field [0001] The invention relates to the field of biology, in particular to protein electrophoresis, and more specifically to the formulation of staining solution for protein electrophoresis strips, and methods for staining and decolorizing using the staining solution. Background technique [0002] Polyacrylamide gel electrophoresis technology is currently the most basic method for analyzing protein components and detection. It can separate proteins and determine their relative molecular weight and protein purity, and can also be used for protein separation and purification. There are many staining methods for polyacrylamide gel electrophoresis, and commonly used methods include amino black staining, Coomassie brilliant blue staining, silver staining, and fluorescent staining. [0003] Among them, Coomassie brilliant blue staining is more sensitive than amino black staining, easier to operate than silver staining, has high repeatability, and has very good mass spe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/30G01N27/447G01N33/68
CPCG01N1/30G01N27/44747G01N33/6803G01N2001/302
Inventor 韦振泉朱佳威任杭洁章丹丹
Owner 浙江玉安康瑞生物科技有限公司
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