Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Kaomas brilliant blue dyeing method and its special gel fixation liquid and dyeing agent

A technology of Coomassie brilliant blue and fixative, which is used in the preparation of test samples, material inspection products, biological tests, etc., can solve problems such as easy breakage, and achieve the effect of avoiding loss, firm protein fixation, and broad application prospects.

Inactive Publication Date: 2007-01-31
INST OF BOTANY CHINESE ACAD OF SCI
View PDF0 Cites 13 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Subsequently, based on the research of Neuhoff et al., Candiano and his collaborators invented a staining method called Blue Silver, which can detect bands containing only 1 nanogram of protein, and the sensitivity is close to that of silver staining. , is said to be the most sensitive one among the reported dyeing methods at present, but this method needs to be decolorized in water for a long time, so that the glue absorbs water and swells, becomes very brittle, and is very easy to break (G.Candiano, M .Bruschi, L.Musante, L.Santucci, G.M.Ghiggeri, B.Carnemolla, P.Orecchia, L.Zardi, P.G.Righetti, Blue silver: A very sensitive colloidal Coomassie G-250 staining for proteome analysis, Electrophoresis 25(2004) 1327 -1333.)

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Kaomas brilliant blue dyeing method and its special gel fixation liquid and dyeing agent
  • Kaomas brilliant blue dyeing method and its special gel fixation liquid and dyeing agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1, silver staining, Blue Silver staining and the comparison of the dyeing effect of the dyeing method of the present invention

[0024] 1. One-dimensional electrophoresis detection of total protein in aerial parts of Salicornia europaea L.

[0025] Gel fixative: an aqueous solution containing 10% acetic acid, 40% ethanol and 10% methanol by volume.

[0026] Coomassie Brilliant Blue Dyeing Mother Solution: Dissolve 2.5 g of Coomassie Brilliant Blue R-250 in 1 liter of ethanol with a concentration of 95% by volume, and shake it in a shaker at 37°C for 3 hours to prepare Coomassie Brilliant Blue Dyeing Mother Solution, store at room temperature , filtered before use.

[0027] Coomassie Brilliant Blue staining agent: use Coomassie Brilliant Blue dyeing mother liquor to prepare an aqueous solution containing 5% acetic acid, 45% ethanol and 0.125% Coomassie Brilliant Blue R-250 in mass volume concentration.

[0028] Decolorization solution I: an aqueous solution ...

Embodiment 2

[0039] Embodiment 2, the sensitivity detection of dyeing method of the present invention

[0040] The detection result of embodiment 1 has initially shown the superiority of the dyeing method of the present invention, now by one-dimensional electrophoresis, the sensitivity of the dyeing method of the present invention is further detected with bovine serum albumin (BSA) as a standard sample, and the loading amount 10μg, 5μg, 3μg, 1μg, 0.5μg, 0.3μg, 0.1μg, 80ng, 50ng, 30ng, 10ng, 8ng and 5ng, the test results are shown in figure 1 Figure D in (Swimming lanes 1-13 are 10 μg, 5 μg, 3 μg, 1 μg, 0.5 μg, 0.3 μg, 0.1 μg, 80ng, 50ng, 30ng, 10ng, 8ng and 5ng of BSA, and lane M is Marker) , when the total protein concentration drops to 10 ng (equivalent to 1 ng / mm 2 ), the protein main band with a molecular weight of about 62kDa in the protein can also be detected (swimming lane 11), indicating that the dyeing method of the present invention has higher sensitivity and can detect protein...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a Coomassie brilliant blue dyeing method and its special gel stationary liquid and coloring agent. It includes the following steps: putting SDS-PAGE glue into the gel stationary liquid for 0.5-2.0h at normal temperature; adding the glue in acetate solution with 0.5-3.0% volume percent concentration to extend for 1.0-3.0h at normal temperature; adding it to the Coomassie brilliant blue coloring agent to dye for 8-24h at normal temperature; add it to decolorizing solution I to decolorize for 1-3h; then adding to decolorizing solution II to decolorize to clear. The invention can detect SDS-PAGE glue protein at nanogram.

Description

technical field [0001] The invention relates to a Coomassie brilliant blue staining method and its special gel fixative and staining agent. Background technique [0002] Although there are many protein staining methods to choose from in proteomics research, the most widely used methods are Coomassie brilliant blue staining (coomassie staining), silver staining and fluorescent staining. Due to the high sensitivity and relatively simple operation procedures, the fluorescent dyes of the SYPRO family have become the most popular dyes (T.H.Steinberg, L.J. Jones, R.P.Haugland, V.L.Singer, SYPRO Orange and SYPRO Red Protein Gel Stains: One-Step Fluorescent Staining of Denaturing Gels for Detection of Nanogram Levels of Protein, Anal.Biochem.239(1996)223-227; M.F.Lopez, K.Berggren, E.Chernokalskaya, A.Lazarev, M.Robinson, W.F.Patton, Acomparison of silver stain and SYPRO Ruby Protein Gel Stain with respect to protein detection in two-dimensional gels and identification by peptide m...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N1/30G01N33/68
Inventor 李银心王旭初
Owner INST OF BOTANY CHINESE ACAD OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com