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Reagent for coloration of protein in gel electrophoresis as well as using method and application

A gel electrophoresis and protein technology, which is applied in the direction of chemical reaction of materials for analysis, chemiluminescence/bioluminescence, etc., to achieve the effect of simple dyeing process, reliable effect and no pollution

Active Publication Date: 2013-12-04
安徽昊拓生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, none of the above methods has substantially changed the problems of cumbersome protein staining steps and serious pollution in gel electrophoresis experiments, and the staining must still be carried out after the electrophoresis is over.

Method used

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  • Reagent for coloration of protein in gel electrophoresis as well as using method and application
  • Reagent for coloration of protein in gel electrophoresis as well as using method and application
  • Reagent for coloration of protein in gel electrophoresis as well as using method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] (1) Solution formula

[0047] 1. Solution A (basic loading buffer without primary amino groups, i.e. boric acid-sodium hydroxide buffer at pH 9.0):

[0048] 20ml 1M boric acid-sodium hydroxide buffer (i.e. Na 2 B 4 o 7 -NaOH)pH9.5

[0049] 4g SDS

[0050] 2g DTT (dithiothreitol)

[0051] 0.1g EDTA (ethylenediaminetetraacetic acid)

[0052] 20ml glycerin

[0053] Add single distilled water to make the volume to 100ml, and the final pH is 9.0.

[0054] 2. Solution B (TRITC solution):

[0055] Dissolve TRITC in DMSO (dimethyl sulfoxide) to prepare a solution with a concentration of 1 mg / ml.

[0056] 3. Coomassie brilliant blue staining solution (for comparison):

[0057] 670ml single distilled water

[0058] 250ml ethanol

[0059] 1.25g Coomassie Brilliant Blue G-250

[0060] After stirring to dissolve, add glacial acetic acid 80ml.

[0061] 4. Decolorization solution (for comparison)

[0062] 670ml single distilled water

[0063] 250ml ethanol

[0064] 80...

Embodiment 2

[0071] (1) Solution formula

[0072] 1. Solution A (basic loading buffer without primary amino groups, i.e. phosphate buffer at pH 8):

[0073] 20ml 1M disodium hydrogen phosphate-sodium dihydrogen phosphate buffer (NaH 2 PO 4 -Na 2 HPO 4 )pH8.0

[0074] 4g SDS

[0075] 2ml β-mercaptoethanol

[0076] 0.1g EDTA

[0077] 20ml glycerin

[0078] The pH was adjusted back to 8 with 3M sodium hydroxide solution.

[0079] Add single distilled water to make up to 100ml.

[0080] 2. Solution B (FITC solution):

[0081] Dissolve FITC in DMF (N,N-dimethylformamide) to prepare a solution with a concentration of 1 mg / ml.

[0082] 3. Coomassie brilliant blue staining solution (for comparison):

[0083] 670ml single distilled water

[0084] 250ml ethanol

[0085] 1.25g Coomassie Brilliant Blue G-250

[0086] After stirring to dissolve, add 80ml of glacial acetic acid.

[0087] 4. Decolorization solution (for comparison)

[0088] 670ml single distilled water

[0089] 250ml et...

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Abstract

The invention discloses a reagent for coloration of protein in gel electrophoresis as well as a using method and application. The reagent comprises a first component and a second component, wherein the first component is the solution formed by dissolving the compound with isothiocyanic acid groups into a solvent for protecting the isothiocyanic acid groups; the second component is the alkaline loading buffer which does not contain amino groups; the using method comprises the steps as follows: mixing the first component and the second component to form treating fluid and mixing the treating fluid with a to-be-tested protein sample, or mixing the second component with the to-be-tested protein sample and then adding the first component, and heating to form the chromoprotein sample solution. According to the invention, the reagent can be used in gel electrophoresis, and has the advantages of simplicity and easiness in use, reliability in effect, low usage amount, easiness in production and processing, wide application range, no pollution, time conservation and labor conversation, capabilities of performing real-time observation and being used for pre-colorating of the protein in gel electrophoresis, and the like, and a protein electrophoresis pattern has no changes which can be observed.

Description

technical field [0001] The invention relates to a protein chromogenic reagent and method, in particular to a reagent for gel electrophoresis protein chromogenic and its usage method and application. Background technique [0002] Gel electrophoresis of proteins is one of the most commonly used experimental methods in life science research. In protein gel electrophoresis experiments, staining agents must be used to stain proteins with eye-catching colors for easy observation of the results. Currently commonly used staining methods such as Coomassie Brilliant Blue staining, silver staining, etc., require cumbersome treatments such as fixing, staining, and decolorizing the gel after the gel electrophoresis of the protein to observe, which takes a long time and makes it impossible to quickly know the experimental results. , and it is not convenient to observe the results during the experiment. Moreover, fixation, staining, and decolorization require a large amount of reagents (...

Claims

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Application Information

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IPC IPC(8): G01N21/76
Inventor 汪弋
Owner 安徽昊拓生物科技有限公司
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