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Commassie brilliant blue staining solution and staining method and application thereof in protein detection

A Coomassie Brilliant Blue and dyeing method technology, which is applied in Coomassie Brilliant Blue staining solution and its dyeing method and in the application field of protein detection, can solve the problem of easy broken glue and other problems, and achieve the effect of not easy to break and low cost

Active Publication Date: 2013-11-27
盛司潼
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The object of the present invention is to provide a kind of Coomassie brilliant blue staining solution and its staining method and the application in protein detection, aim at guaranteeing sensitivity (10ng) under the situation, further reduce SDS-PAGE gel dyeing cost, and solve the problem of gel staining after decolorization. easily broken problem

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  • Commassie brilliant blue staining solution and staining method and application thereof in protein detection
  • Commassie brilliant blue staining solution and staining method and application thereof in protein detection
  • Commassie brilliant blue staining solution and staining method and application thereof in protein detection

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Embodiment Construction

[0030] In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments.

[0031] A kind of Coomassie brilliant blue staining solution, described staining solution is to comprise the alcohol of 10%~20% in volume percent concentration (v / v), and the acid of 4.5%~10% in volume percent concentration (v / v), The mass volume concentration (w / v) is the ammonium sulfate of 5%~10%, and the mass volume concentration (w / v) is the aqueous solution of Coomassie brilliant blue of 0.01%~0.1%; The alcohol comprises at least one in methanol and ethanol species; the acid includes at least one of phosphoric acid and acetic acid; the Coomassie Brilliant Blue includes at least one of G250 and R250.

[0032] The higher the concentration of Coomassie Brilliant Blue, the deeper the variegation of the protein in the SD-PAGE gel, that is, the high...

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Abstract

The invention relates to the field of protein staining, in particular to a Commassie brilliant blue staining solution and a staining method and application of the Commassie brilliant blue staining solution in protein detection. The Commassie brilliant blue staining solution is an aqueous solution containing 10-20% (v / v) of alcohol, 4.5-10% (v / v) of acid, 5-10% (w / v) of ammonium sulfate and 0.01-0.1% (w / v) of Commassie brilliant blue. The staining method comprises the following steps: (A) staining: putting SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) gel in the Commassie brilliant blue staining solution and staining for more than 45min; and (B) destaining: flushing the stained SDS-PAGE gel with water till the staining solution is completely washed off. Through adoption of the Commassie brilliant blue staining solution and the staining method provided by the invention, under the condition that the sensitivity is not reduced, the SDS-PAGE gel is stained at low cost; moreover, the detained gel is hardly broken, thereby brining no trouble to the following mass spectrum research and the like.

Description

technical field [0001] The invention relates to the field of protein staining, more specifically, to a Coomassie brilliant blue staining solution, a staining method and an application in protein detection. Background technique [0002] At present, there are roughly four protein staining methods in proteomics research, Coomassie brilliant blue staining, fluorescent staining, silver nitrate staining, and isotope colorimetry. Among them, fluorescent staining is mainly based on SYPRO reagent, which has high protein detection sensitivity and is compatible with mass spectrometry, but it has not been used as a routine method due to the need for special detection instruments and expensive reagents; isotope colorimetry has safety and operational limitations However, silver nitrate staining is currently recognized as the most sensitive protein detection method except for radioactive labeling, and can detect proteins on SDS-PAGE gels at the nanogram level. However, The steps are cumbe...

Claims

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Application Information

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IPC IPC(8): C09B67/14G01N1/30
Inventor 盛司潼
Owner 盛司潼
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