Show Order Info
SKU AR0140
Pack size1kit (for 10 assays to stain gels of 5 x 8.5cm)
FormLiquid
ApplicationsFor protein staining on polyacrylamide gels.

List of Components

DescriptionQuantityVolumeContentsCatalog Number
Commassie Blue Staining Solution1100mL0.1% (w/v) Brilliant Blue R-250, 25% (v/v) methanol and 5% (v/v) acetic acidAR0140-1
Commassie Blue Destaining Solution1100mL25% (v/v) methanol and 5% (v/v) acetic acidAR0140-2

Product Overview

Product Name Commassie Blue Staining Destaining Solution
SKU/Catalog Number AR0140
Form Liquid
Size 1kit (for 10 assays to stain gels of 5 x 8.5cm)
Contents 0.1% (w/v) Brilliant Blue R-250, 25% (v/v) methanol and 5% (v/v) acetic acid.
Working Concentration 10mL for each gel
Storage Upon receipt store at room temperature. It is stable at room temperature for one year.
Target Molecule Protein
Label or Dye Coomassie Blue R-250
Sensitivity 10ng
Equivalent Millipore Sigma (Product No. B8522)
Application For protein staining on polyacrylamide gels.
Compatibility Compatible with mass spectrometric analysis
Cite This Product Commassie Blue Staining Destaining Solution (Boster Biological Technology, Pleasanton CA, USA, Catalog # AR0140)

Introduction

Usually after electrophoresis, researchers should evaluate the result through staining and destaining, and detect the protein which was disassembled into different bands from the gel. Besides, researchers also could electroblotting the gel for different research purpose. This solution is a coomassie blue R-250 dye-based, can be widely used for staining of protein electrophoresis gel of SDS-PAGE or non-denaturing PAGE, as well as in the residual protein detection of PAGE gel after by Western transferring. By this means, the working range for protein electrophoresis bands can be as low as 10ng. This production is acidic and its corrosion requires a necessary protection when use it.

Features

• Easy detection—Develops intensely colored complexes with proteins
• High sensitivity—Can determine as little as 10 ng of protein present in a gel matrix
• Reversible staining—Anion of Coomassie Brilliant Blue formed in the acidic staining medium combines with the protonated amino groups of proteins by electrostatic interaction; resulting complex is reversible under the proper conditions
• Differentiation between bound and unbound dye—When dissolved in 0.01M citrate buffer at pH 3.0, has an absorption maximum at 555nm; protein-dye complex is characterized by a peak slightly broader than that of the free dye with a maximum at 549 nm

Background

Coomassie Brilliant Blue R-250 is one of the most common forms of coomassie dye, which is a key component of various colorimetric protein gel stains. Coomassie R-250 and G-250 dyes are two chemical forms of a disulfonated triphenylmethane compound that is commonly used as the basis of stains for detection of proteins in gel electrophoresis and Bradford-type assay reagents for protein quantitation. The R-250 (red-tinted) form lacks two methyl groups that are present in the G-250 (green-tinted) form, which is also called colloidal coomassie dye. Typically, coomassie gel stains and protein assay reagents are formulated as very acidic solutions in 25 to 50% methanol. In acidic conditions, the dye binds to proteins primarily through basic amino acids (primarily arginine, lysine and histidine), and the number of coomassie dye ligands bound to each protein molecule is approximately proportional to the number of positive charges found on the protein. Protein-binding causes the dye to change from reddish-brown to bright blue (absorption maximum equals 595 nm).

Protocol

1. After electrophoresis, place gel in a clean tray. Wash the gel with 100mL of distilled water 3 times for 5-10 min each with gentle agitation. This step is to gently remove the interference of SDS and salt.

2. Discard last wash. Add sufficient volume of Commassie Blue Staining Solution to cover the gel.
Note: Shake the Commassie Blue Staining Solution before use.

3. Incubate at room temperature for 60 minutes with gentle agitation.

4. Discard the staining solution and wash the gel for one hour or overnight at room temperature with Commassie Blue Destaining Solution.

Note: For the gel thicker than 1.5mm, it takes a longer period for above mentioned washing and staining steps.

Troubleshooting

ProblemPossible CauseSolution
No band is seenSample is not enoughTry to add two BSA lanes with different volumes when electrophoresis as a positive control.
High backgroundImpurity is not removed completelyTry to wash longer or wash more times
Precipitate occurs when staining the gelthe container was pollutedTry to use a clean container while using new stain solution until the bands are seen.

Coomassie Blue Staining Destaining Solution Images

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Coomassie Blue Staining Destaining Solution
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