SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) coomassie brilliant blue R250 fast staining solution, staining method and application

A Coomassie brilliant blue, rapid dyeing technology, applied in the preparation of test samples, material inspection products, measuring devices, etc., can solve the problems of increasing the probability of sample contamination, prolonging the operation time, cumbersome fixing and sensitization steps, etc. The effect of shortening the dyeing process time, background uniformity, and shortening of the dyeing time

Inactive Publication Date: 2012-07-25
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, the steps of fixation and sensitization before staining are cumbersome, whi

Method used

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  • SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) coomassie brilliant blue R250 fast staining solution, staining method and application
  • SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) coomassie brilliant blue R250 fast staining solution, staining method and application
  • SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) coomassie brilliant blue R250 fast staining solution, staining method and application

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0033] Example 1

[0034] (1) The fast dyeing solution used in this embodiment:

[0035] Components with the following content: ethanol with a volume percentage of 30%, methanol with a volume percentage of 10%, CBB R250 with a mass concentration of 1.25g / L, phosphoric acid with a volume percentage of 5%, mass concentration It is 100g / L of ammonium sulfate.

[0036] Specifically formulated in the following ways:

[0037] Staining solution 11: Measure 300mL ethanol, 100mL methanol, and electronic balance to weigh 1.25g CBB R250 powder in pure water, and dissolve it to 500mL with pure water. After mixing, seal it and place it overnight at 37°C at 100 revolutions / min. The dye is fully dissolved.

[0038] Weigh 100 g of ammonium sulfate on the electronic balance of dyeing solution 21, add 300 mL of pure water to fully dissolve it, measure 59 mL of 85% phosphoric acid in a graduated cylinder, and dilute the pure water to 500 mL.

[0039] Mix the staining solution 11 and the staining solution...

Example Embodiment

[0055] Example 2

[0056] (1) The fast dyeing solution used in this embodiment:

[0057] Components with the following content: ethanol with a volume percentage of 30%, methanol with a volume percentage of 10%, CBB R250 with a mass concentration of 1.25g / L, phosphoric acid with a volume percentage of 5%, mass concentration It is 100g / L of ammonium sulfate.

[0058] The preparation process is the same as in Example 1.

[0059] (2) Extraction of C whey protein from Hevea brasiliensis:

[0060] The C whey protein of Hevea brasiliensis was extracted by the BPP method (the extraction method is the same as above). The fresh latex collected was ultracentrifugated to separate the C whey component, dissolved in an equal volume of protein lysate BPP, and Tris balanced phenol was used for protein extraction For extraction, supersaturated ammonium sulfate is used to precipitate the protein, and the lysis solution is used to lyse the protein precipitation to obtain a protein solution. The Bradfor...

Example Embodiment

[0075] Example 3

[0076] (1) The components and preparation method of the fast dyeing solution used in this embodiment are the same as those in embodiment 1, and the dyeing solution R is evenly divided into two equal parts;

[0077] (2) One-way SDS-PAGE electrophoresis of cassava leaf protein, Hevea C whey protein and bovine serum protein:

[0078] Use SDS-PAGE with a mass percentage of 12.5% ​​to prepare two separation gels with a thickness of 1.5 mm. After gelling, prepare a concentrated SDS-PAGE gel with a mass percentage of 4%. Use a 15-tooth comb to make 15 holes Concentrated gel, the maximum sample volume is 100 microliters. Add 30 micrograms of the protein solution to the sample hole of the gel in the order of cassava leaf protein, Hevea C whey protein, and bovine serum protein. Use Hoefer SE 600chroma one-way gel electrophoresis instrument to set constant power parameters , Perform one-way SDS-PAGE electrophoresis.

[0079] (3) Staining and decolorization of gel:

[0080] Tw...

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Abstract

The invention discloses SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) coomassie brilliant blue R250 fast staining solution, which includes from 20 to 40% of ethanol by volume percentage, from 5 to 10% of carbinol by volume percentage, from 0.5 to 1.25g/L of CBB (coomassie brilliant blue) R250 by mass concentration, from 2.5 to 5% of phosphoric acid by volume percentage and from 100 to 200g/L of ammonium sulfate by mass concentration. A fixing step and a sensitization step before staining are omitted, protein staining time can be shortened, and the possibility that protein samples are polluted is greatly reduced. The invention further discloses a method for staining SDS-PAGE gel by the aid of the staining solution and application of the staining solution in a protein two-dimensional gel electrophoresis and protein western blot qualitative diagnosis and/or quantification process.

Description

technical field [0001] The present invention relates to a Coomassie Brilliant Blue fast staining solution, a dyeing method and an application thereof, in particular to a SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel) Coomassie Brilliant Blue R250 fast staining solution, a dyeing method and an application . Background technique [0002] Proteomics research can reveal the laws of life by constructing a map of temporal and spatial differences in protein expression in cells. The SDS polyacrylamide gel electrophoresis separation technology of protein is the key technology of proteomics research, and the staining of the gel is the core technology to obtain the analyzable map. In protein analysis and detection, Coomassie brilliant blue staining, silver nitrate staining and fluorescent dye staining are three staining methods widely used in protein analysis and detection. The silver nitrate staining method is recognized as a gel staining method with high sensitivity, and the ...

Claims

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Application Information

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IPC IPC(8): G01N1/30G01N27/447G01N33/68
Inventor 王旭初王海燕郭安平王丹常丽丽
Owner INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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