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A kind of Coomassie Brilliant Blue G250 dyeing method and its special dyeing solution and application

A technology of Coomassie brilliant blue and CBBG-250, applied in the preparation of test samples, etc., can solve the problems of cumbersome steps, limited application scope, long time consumption, etc., and achieve the effect of simplifying dyeing steps, improving sensitivity and wide application range.

Inactive Publication Date: 2011-12-21
INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this staining method is cumbersome and time-consuming, which limits its application

Method used

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  • A kind of Coomassie Brilliant Blue G250 dyeing method and its special dyeing solution and application
  • A kind of Coomassie Brilliant Blue G250 dyeing method and its special dyeing solution and application
  • A kind of Coomassie Brilliant Blue G250 dyeing method and its special dyeing solution and application

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Experimental program
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Effect test

Embodiment 1

[0033] The Coomassie Brilliant Blue staining solution provided by this implementation contains the following components: CBB G-250 with a mass volume concentration of 0.125%, ammonium sulfate with a mass volume concentration of 10%, phosphoric acid with a volume percentage of 5%, and a volume percentage of 30% ethanol and 10% methanol by volume, each solution uses ddH2 O preparation, its specific preparation process is as follows:

[0034] (1) Mix 300mL ethanol, 100mL methanol and 100mL ddH 2 O mixed together;

[0035] (2) Weigh 1.25g CBB G-250, and add it into the mixed solution of (1);

[0036] (3) Put the solution obtained in (2) on a shaker, set the speed at 150rpm, and shake at 37°C for more than one hour to ensure that CBBG-250 is completely dissolved, then put it at room temperature for later use, and call this solution CBB-A liquid;

[0037] (4) Weigh 100g ammonium sulfate, add 300mLddH 2 O, until ammonium sulfate is completely dissolved;

[0038] (5) Add 58.8mL o...

Embodiment 2

[0078]The Coomassie Brilliant Blue staining solution provided by this implementation contains the following components: CBB G-250 with a mass volume concentration of 0.125%, ammonium sulfate with a mass volume concentration of 10%, phosphoric acid with a volume percentage of 5%, and a volume percentage of 30% ethanol and 10% methanol by volume, each solution uses ddH 2 O preparation, its specific preparation process is as follows:

[0079] (1) Mix 300mL ethanol, 100mL methanol and 100mL ddH 2 O mixed together;

[0080] (2) Weigh 1.25g CBB G-250, and add it into the mixed solution of (1);

[0081] (3) Put the solution obtained in (2) on a shaker, set the speed at 150rpm, and shake at 37°C for more than one hour to ensure that CBBG-250 is completely dissolved, then put it at room temperature for later use, and call this solution CBB-A liquid;

[0082] (4) Weigh 100g ammonium sulfate, add 300mLddH 2 O, until ammonium sulfate is completely dissolved;

[0083] (5) Add 58.8mL ...

Embodiment 3

[0100] The Coomassie Brilliant Blue staining solution provided by this implementation contains the following components: CBB G-250 with a mass volume concentration of 0.10%, ammonium sulfate with a mass volume concentration of 15%, phosphoric acid with a volume percentage of 3%, and a volume percentage of 40% ethanol and 5% methanol by volume, each solution uses ddH 2 O preparation, its specific preparation process is as follows:

[0101] (1) Mix 400mL ethanol, 50mL methanol and 100mL ddH 2 O mixed together;

[0102] (2) Weigh 1.0g CBB G-250, and add it into the mixed solution of (1);

[0103] (3) Put the solution obtained in (2) on a shaker, set the speed at 150rpm, and shake at 37°C for more than one hour to ensure that CBBG-250 is completely dissolved, then put it at room temperature for later use, and call this solution CBB-A liquid;

[0104] (4) Weigh 150g ammonium sulfate, add 300mLddH 2 O, until ammonium sulfate is completely dissolved;

[0105] (5) Add 35.3mL of ...

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Abstract

The invention discloses a Coomassie Brilliant Blue G250 staining liquid, which contains the following components: CBB G-250 with a mass volume concentration of 0.1-0.2%, ammonium sulfate with a mass volume concentration of 5-15%, and a volume percentage of 3 -8% phosphoric acid, 20-40% ethanol by volume, and 5-20% methanol by volume. The staining solution integrates the fixative solution, the sensitizing solution and the staining solution, the background of the dyed gel is lighter, and the dyeing sensitivity is high; a method for dyeing by using the above-mentioned Coomassie Brilliant Blue G250 staining solution is also disclosed. In addition to high repeatability and low staining background, this detection method is fast and sensitive, and the number of detected protein spots is relatively large; it has good mass spectrometry compatibility and can reduce protein modification in vitro; it has a wide range of applications. At the same time, the application of the above method in the qualitative and / or quantitative detection of proteins is disclosed.

Description

technical field [0001] The invention relates to a dyeing method of Coomassie Brilliant Blue and its dyeing solution and application, in particular to a Coomassie Brilliant Blue G250 dyeing method and its special dyeing solution and application. Background technique [0002] Two-dimensional electrophoresis and mass spectrometry (MS) are the two most important technologies in proteomics research. After SDS-PAGE, sensitive and efficient protein staining becomes a prerequisite for subsequent proteomics research (Choi et al.1996; Wang et al. .2007a; Yasumitsu et al.2010b). In protein analysis and detection, cox staining, silver staining and fluorescent staining are three staining methods widely used in protein analysis and detection (Patton, 2000). Among these methods, silver staining is currently recognized as the most sensitive protein detection method except radioactive labeling, which can detect proteins at the nanogram level (Yan et al.2000; Candiano et al.2004; Yasumitsu e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/30
Inventor 王旭初郭安平王海燕王东阳
Owner INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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