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High sensitivity and low background Coomassie brilliant blue staining solution and using method thereof

A technology of Coomassie brilliant blue and high sensitivity is applied in the field of high sensitivity and low background Coomassie brilliant blue staining solution, which can solve the problems of poor expression of low-abundance proteins and low staining sensitivity, so as to shorten detection time, improve detection sensitivity and improve work efficiency. The effect of efficiency

Inactive Publication Date: 2019-07-30
湖北擎科生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the main problem of CBB R250 is that the staining sensitivity is not high, and the visualization of low-abundance proteins is poor. Many studies have made efforts to reduce its staining background and improve its detection sensitivity. There are many domestic and foreign researches on CBB staining methods. reported, but most of them take hours or days to complete the staining and destaining process, and rarely report staining sensitivity at the nanogram level

Method used

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  • High sensitivity and low background Coomassie brilliant blue staining solution and using method thereof
  • High sensitivity and low background Coomassie brilliant blue staining solution and using method thereof
  • High sensitivity and low background Coomassie brilliant blue staining solution and using method thereof

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Embodiment 1

[0025] The high-sensitivity and low-background Coomassie Brilliant Blue staining solution provided by the invention comprises: 0.03% CBB R250 (w / v), 14% ethanol (v / v), 10% ammonium sulfate (w / v), 4% acetic acid ( v / v).

[0026] Protein sample preparation: the protein sample is BSA (Albumin from bovine serum), dissolve the sample with deionized water, and dilute it into different concentrations. 15.6ng, 7.8ng, 3.9ng, 1.9ng, 1ng, 0.5ng.

[0027] Staining process: After electrophoresis, take out the gel, wash it with sufficient deionized water for 2-3 times, each time for 5 minutes, put the gel in an appropriate amount of prepared staining solution, shake and stain on the decolorization shaker for 10 minutes And 1h, after the staining, shake and wash the gel with deionized water for 1-5min, take out the gel and scan to observe the staining effect.

[0028] The staining result is as figure 1 As shown, the minimum detection limit can reach 7.8ng after 10 minutes of staining with...

Embodiment 2

[0030] The high-sensitivity and low-background Coomassie Brilliant Blue staining solution provided by the invention comprises: 0.03% CBBR250 (w / v), 12% ethanol (v / v), 8% ammonium sulfate (w / v), 2% acetic acid (v / v).

[0031] Protein sample preparation was the same as in Example 1.

[0032] The dyeing process is the same as in Example 1.

[0033] The staining result is as figure 1 As shown, the minimum detection limit can reach 15.6 ng after 10 minutes of staining with the dye of the present invention, and after 1 hour of staining, the minimum detection limit can reach 7.8 ng, and the background is clear.

Embodiment 3

[0035] The high-sensitivity and low-background Coomassie Brilliant Blue staining solution provided by the invention comprises: 0.05% CBBR250 (w / v), 16% ethanol (v / v), 15% ammonium sulfate (w / v), 6% acetic acid (v / v).

[0036] Protein sample preparation was the same as in Example 1.

[0037] The dyeing process is the same as in Example 1.

[0038] The staining result is as image 3 As shown, the minimum detection limit can reach 7.8ng after 10 minutes of staining with the dyeing agent of the present invention, and after 1 hour of staining, the minimum detection limit can reach 3.9ng, and the background is clear.

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Abstract

The present invention discloses a high sensitivity and low background Coomassie brilliant blue (CBB) staining solution and a using method thereof, and belongs to the technical field of protein staining reagents of SDS-PAGE. The staining solution is composed of CBB R250 with a mass volume concentration of 0.03%-0.05%, acetic acid with a volume percentage of 2%-6%, ammonium sulfate with a mass volume concentration of 8%-15%, and ethanol with a volume percentage of 12%-16%. The CBB staining solution provided by the present invention is optimized on the basis of the traditional staining solution,and a clearer background is obtained, so that the detection sensitivity is greatly improved and the staining sensitivity reaches the nanogram level. According to the method for using the CBB stainingsolution provided by the present invention, without elution after staining, deionized water is used to wash to destroy the colloid state to complete decolorization, so that the detection time is greatly shortened, the method can be used for rapid protein staining of SDS-PAGE, and work efficiency can be improved.

Description

technical field [0001] The invention relates to the technical field of protein staining reagents for SDS-PAGE, in particular to a high-sensitivity and low-background Coomassie brilliant blue staining solution and a method for using the same. Background technique [0002] Protein band staining is an indispensable key technical link in SDS-PAGE. At present, the staining methods after protein electrophoresis mainly include Coomassie brilliant blue staining, silver staining, and fluorescent staining. Silver staining has high sensitivity, but is often incompatible with mass spectrometry. Fluorescent staining has high sensitivity and is compatible with mass spectrometry, but requires special detection instruments, and is generally not used as a routine method. [0003] CBB (Coomassie Brilliant Blue) staining is relatively inexpensive, reproducible, and does not require expensive equipment, and is also the most popular. CBB includes CBB R250 and CBB G250, the former is the most co...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/30
CPCG01N1/30
Inventor 伍潇洁
Owner 湖北擎科生物科技有限公司
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