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Silver staining reagent and staining method thereof

A technology for reagents and staining solutions, applied in the biological field, can solve the problems of inability to display protein components, low accuracy, low sensitivity, etc., and achieve the effect of shortening the time required for detection, accurate detection, and improving sensitivity

Inactive Publication Date: 2012-06-20
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This staining method has low sensitivity, and some trace protein components in the sample are often not displayed, which is time-consuming and has low accuracy.
In order to improve the sensitivity of protein detection, silver staining method can also be used, but the existing silver staining method steps are cumbersome and time-consuming

Method used

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  • Silver staining reagent and staining method thereof
  • Silver staining reagent and staining method thereof
  • Silver staining reagent and staining method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1, silver staining staining of protein

[0035] 1. Protein electrophoresis

[0036] Equipment used in electrophoresis: electrophoresis instrument Bio-Rad mini protean III vertical plate electrophoresis system

[0037] 1. Preparation of SDS-PAGE electrophoresis gel: the concentration of the separating gel is 15%, the concentration of the stacking gel is 4.5%, and the thickness of the gel is 0.75mm.

[0038] Table 1 Preparation of SDS-PAGE electrophoresis gel

[0039]

[0040] in:

[0041] (1) 30% acrylamide (w / v): 29g acrylamide; 1g N,N'methylenebisacrylamide; dilute to 100mL with deionized water, filter and store.

[0042] (2) Lower gel buffer solution (pH 8.8): 18.17g Tris; 0.4g SDS dissolved in deionized water, adjust the pH to 8.8 with HCl, and dilute to 100mL.

[0043] (3) Supergel buffer solution (pH 6.8): 12.1g Tris; 0.4g SDS was dissolved in deionized water, and HCl was added to adjust the pH to 6.8, and the volume was adjusted to 100mL.

[004...

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Abstract

The invention discloses a silver staining reagent and a staining method thereof. The invention provides a reagent for protein silver staining. The reagent provided by the invention comprises a staining liquid and a developing liquid; the solute of the straining liquid in the reagent comprises the following substances of ethanol, nitric acid and silver nitrate, wherein the proportion of the ethanol to the nitric acid to the silver nitrate is 25 ml: 1 ml: 0.2 g; and the developing liquid in the reagent comprises the following substances of sodium carbonate and formaldehyde, wherein the proportion of the sodium carbonate to the formaldehyde is 3 g: 0.2 ml. Proved by experiments, a novel silver staining method and a specific silver staining reagent are adopted in the invention, so that the operation is simple, the sensitivity is largely improved, and rapid, high-efficient and accurate detection of protein is realized after polyacrylamide gel electrophoresis.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a silver staining reagent and a staining method thereof. Background technique [0002] Polyacrylamide gel electrophoresis is one of the commonly used protein detection techniques in protein research. Protein staining is a routine technique for detecting protein bands on electrophoretic gels and a way to visualize proteins. Silver staining is one of the most sensitive determination methods of protein gel electrophoresis, but the steps are cumbersome, requiring more preparation reagents, and the cost is high. [0003] Proteins are usually stained with Coomassie brilliant blue after polyacrylamide gel electrophoresis (PAGE). This staining method has low sensitivity, and some trace protein components in the sample are often not displayed, which is time-consuming and has low accuracy. In order to improve the sensitivity of protein detection, silver staining method can also be used, but...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/30
Inventor 班丽萍赵立静石旺鹏
Owner CHINA AGRI UNIV
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