Low toxicity low irritation Commassie brilliant blue fast staining solution and dyeing and decoloring method

A Coomassie brilliant blue, fast dyeing technology, applied in the field of molecular biology technology and proteomics, can solve the problems of pollution, high sensitivity and high cost of dyeing reagents in the dyeing process, achieve dyeing resolution and high color fastness, toxic and irritating The effect of low sexual components and shortened dyeing and decolorization time

Active Publication Date: 2019-04-16
苏州译酶生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, gel preparation and electrophoresis have mature technology and special equipment. At present, the products commonly used in the market for gel staining and decolorization mainly have the following defects: 1) the sensitivity and resolution are low, and the decolorization time is not easy to control. The results of gel staining after decolorization are not easy to preserve; 2) the silver nitrate staining method with higher sensitivity has complicated staining steps, the staining process is more polluted, and the cost of reagent raw materials used in staining is higher; 3) the fluorescent staining method and negative staini

Method used

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  • Low toxicity low irritation Commassie brilliant blue fast staining solution and dyeing and decoloring method
  • Low toxicity low irritation Commassie brilliant blue fast staining solution and dyeing and decoloring method
  • Low toxicity low irritation Commassie brilliant blue fast staining solution and dyeing and decoloring method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 Determination of Coomassie Brilliant Blue G250 Concentration in Coomassie Brilliant Blue Rapid Staining Solution

[0048] Coomassie brilliant blue is divided into R-150, R-250, R-350, G-250 and so on. Among them, R-350 is the most sensitive, R is red-blue, and G is green-blue. R-250 is triphenylmethane, each molecule contains two SO 3 The H group is acidic, and like amino black, it also binds to the basic group of the protein. G-250 is dimethycyanine brilliant blue, which is a kind of triphenylmethane substituted by methyl group. The present invention replaces Coomassie brilliant blue R-250, which is slower to dye, with faster dyeing method on the basis of traditional Coomassie brilliant blue dyeing method. The fast Coomassie Brilliant Blue G-250 effectively shortens the staining time during the staining process. When Coomassie Brilliant Blue G-250 is at a certain pH, the colored complex formed by this dye and protein can be completely depolymerized, and it...

Embodiment 2

[0055] Example 2 Effects of adding different alcohols on the staining results in Coomassie Brilliant Blue Rapid Staining Solution

[0056] Alcohols play a role in fixing proteins in the staining solution. Adding alcohols and other organic components can improve the signal-to-noise ratio of gel staining, make the background of the staining results clearer, and reduce the concentration of some residual substances on the gel surface during the staining process. interference. Although all alcohol components in the dyeing solution are removed, the dyeing solution can also guarantee the same rapid dyeing effect, but the time for dissolving the dye during the preparation process will be extended from 5-10 minutes to 60-90 minutes, and the prepared dyeing solution should be stored at room temperature for 1- After 2 weeks, a large amount of dye precipitation will appear at the bottom, and the dyeing time needs to be extended to achieve the same dyeing effect as before. The unused alco...

Embodiment 3

[0064] Determination of Copper Sulfate / Chromium Trichloride Concentration in Coomassie Brilliant Blue Rapid Staining Solution of Example 3

[0065] The product used in this example for fusion expression in Escherichia coli has a certain endonuclease with a 6×His tag, and its size is about 78 kDa. Escherichia coli induced expression disruption solution, after DNase I treatment, use Perform Heparin affinity chromatography purification, and take collected protein samples at different stages for SDS-PAGE.

[0066] The three formulations of the Coomassie Brilliant Blue Quick Staining Solution with a volume of 1 L containing different concentrations of copper sulfate are as follows:

[0067]

[0068]

[0069]

[0070] As shown in Figure 3(a)-(c), there is no significant difference in the staining time of Coomassie brilliant blue rapid staining solution containing different concentrations of anhydrous copper sulfate, and the staining time of Coomassie brilliant blue rapid ...

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Abstract

The invention discloses a low toxicity low irritation Commassie brilliant blue fast staining solution and a dyeing and decoloring method. The low toxicity low irritation Commassie brilliant blue faststaining solution contains 0.005 percent-0.02 percent of Commassie brilliant blue, 1 percent -5 percent (v/v) of concentrated hydrochloric acid or 1 percent -5 percent (v/v) of 85% phosphate, 1 percent -5 percent (v/v) of glacial acetic acid, 1.5 per thousand-5 per thousand of anhydrous copper sulfate or chromium trichloride, and 3 percent -5 percent (v/v) of mixed water solution of absolute ethanol. The staining solution has low toxicity and low irritation, dyeing and decoloring time is short, and dyeing resolution and color fastness are high.

Description

technical field [0001] The invention relates to the technical fields of molecular biology technology and proteology, and mainly relates to a Coomassie brilliant blue rapid staining solution and a staining and decolorization method applied to protein polyacrylamide gel electrophoresis (SDS-PAGE). Background technique [0002] Polyacrylamide gel electrophoresis (sodium dodecyl sulfate polyacrylamide gelelectrophoresis, SDS-PAGE) has become an important technical means in modern biological research, and it is an economical, rapid and reproducible method for identifying the purity and content of proteins. SDS-PAGE electrophoresis is easy to operate and has a wide range of uses. It has become an important analytical technique in many research fields. SDS-PAGE includes four main steps, namely: gel preparation, electrophoresis, staining, and decolorization. Among them, gel preparation and electrophoresis have mature technology and special equipment. At present, the products commonl...

Claims

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Application Information

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IPC IPC(8): G01N1/30G01N27/447G01N33/68
CPCG01N1/30G01N27/447G01N33/68
Inventor 张惠丹戴敬杨晟姜坤廷李莹玉
Owner 苏州译酶生物科技有限公司
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