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High-sensitivity rapid SDS-PAGE albumen glue staining solution

A high-sensitivity, dyeing solution technology, applied in the preparation of SDS-PAGE protein glue dyeing solution, can solve the problems of high toxicity of dyeing solution and decolorizing solution, volatilization of irritating gas, harm to laboratory personnel, etc., to achieve low toxicity and avoid pollution. , the effect of high sensitivity

Inactive Publication Date: 2020-07-31
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Polyacrylamide gel electrophoresis is a technique widely used in molecular biology experiments. The color development of protein gel is a very critical step. The traditional color development method is mainly divided into two steps: staining and decolorization. The dyeing solution and decolorization solution used in this process are highly toxic, and it is easy to cause the volatilization of irritating gas during the operation, which brings harm to the experimenters, and the decolorization process needs to be repeated, which is very cumbersome.
In recent years, a protein glue staining instrument newly developed can quickly stain protein glue and is easy to operate, but the cost is very expensive in the process of use, and most laboratories cannot afford it, so it has faded out of the market

Method used

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  • High-sensitivity rapid SDS-PAGE albumen glue staining solution
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  • High-sensitivity rapid SDS-PAGE albumen glue staining solution

Examples

Experimental program
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Embodiment 1

[0017] The embodiment of the invention discloses a high-sensitivity rapid protein glue staining solution, which comprises the following components: Coomassie Brilliant Blue G-25050mg, absolute ethanol 50ml, phosphoric acid 50ml, dextrin 7.5g, purified water 400ml.

[0018] At the same time, the present invention and the traditional CBB G-250 staining solution were used to stain two pieces of protein SDS-PAGE with the same loading amount, and the staining results and sensitivities were compared.

[0019] Protein sample loading: 1mg / ml, 0.5mg / ml, 0.1mg / ml, 0.05mg / ml, 0.01mg / ml were prepared by polyacrylamide gel electrophoresis with bovine serum albumin (BSA) as the standard sample , 0.005 mg / ml BSA solution. The loading volume was 6 μl. The final loading amounts were 6μg, 3μg, 0.6μg, 0.3μg, 60ng, and 30ng, respectively.

[0020] Staining process of protein polyacrylamide gel: Pour 30ml each of the staining solution of the present invention and CBB G-250 staining solution into...

Embodiment 2

[0023] The staining solution provided in the embodiment of the present invention includes the following components: 50 mg of Coomassie Brilliant Blue G-250, 50 ml of absolute ethanol, 50 ml of phosphoric acid, 5 g of dextrin, and 400 ml of purified water.

[0024] Protein sample loading: through polyacrylamide gel electrophoresis, bovine serum albumin (BSA) was used as a standard sample to prepare 1 mg / ml, 0.1 mg / ml, and 0.01 mg / ml BSA solutions. The loading volume was 6 μl. The final loading amounts were 6 μg, 0.6 μg, and 60 ng, respectively.

[0025] The dyeing process is the same as in Example 1.

[0026] Experimental results: the dyeing effect of the staining solution of the present invention on protein SDS-PAGE is as attached image 3 As shown, the protein SDS-PAGE band stained with the staining solution added with 5g dextrin is clear, the background is clean, and the sensitivity is high.

Embodiment 3

[0028] The staining solution provided in the embodiment of the present invention includes the following components: 50 mg of Coomassie Brilliant Blue G-250, 50 ml of absolute ethanol, 50 ml of phosphoric acid, 1 g of dextrin, and 400 ml of purified water.

[0029] Protein sample loading process is the same as in Example 2

[0030] The dyeing process is the same as in Example 1.

[0031] Experimental results: the dyeing effect of the staining solution of the present invention on protein SDS-PAGE is as attached Figure 4 As shown, only 1g of dextrin was added to the staining solution to stain weak protein SDS-PAGE bands, and the sensitivity was relatively low.

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Abstract

The invention relates to a high-sensitivity rapid SDS-PAGE albumen glue staining solution. Every 500 ml of staining solution is prepared from the following materials: 5-10 g of dextrin; 25-75 mg of Coomassie brilliant blue G-250; 40-60 ml of absolute ethyl alcohol; 40-60 ml of phosphoric acid; remaining purified water. The staining solution has low toxicity and can effectively reduce the harm to operators in the preparation and use processes and avoid pollution to the experimental environment; the operation process is very simple, a strip can be seen only by soaking albumen glue in a stainingsolution and then placing the staining solution on a shaking table for staining, and fixation and decoloration treatment are not needed; the sensitivity is high, and 30 ng of protein strips in the gelcan be distinguished after dyeing for 20 minutes; the reaction is rapid, and when the protein amount reaches 6mu g, the strip can be shown in only one minute.

Description

technical field [0001] The technical scheme of the present invention relates to the field of molecular biology experiments, in particular to a preparation method of SDS-PAGE protein glue staining solution. Background technique [0002] Polyacrylamide gel electrophoresis is a technique widely used in molecular biology experiments. The color development of protein gel is a very critical step. The traditional color development method is mainly divided into two steps: staining and decolorization. The dyeing solution and decolorization solution used in this process are highly toxic, and the irritating gas is easily volatilized during the operation, which brings harm to the experimenters. In addition, repeated decolorization is required, and the process is very cumbersome. In recent years, a newly developed protein glue staining instrument can quickly stain protein glue and is easy to operate, but the cost is very expensive in the process of use, and most laboratories cannot affor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/30
CPCG01N1/30G01N2001/302
Inventor 王鸣飞许英健杨雪沈月全
Owner NANKAI UNIV