What does a Coomassie stain do?
Coomassie Blue stain is used to stain the protein bands in polyacrylamide gels. One common way to use it is to dissolve the dye in a mixture of methanol, acetic acid, and water. This stain will permeate the gel, stain the protein, and also fix the protein in place.
Can you Coomassie stain overnight?
After incubation, discard the stain. Stain cannot be re-used. Note: Gel can be stained for up to 3 hours, but after 3 hours, sensitivity will decrease. If you need to leave the gel overnight in the stain, add 2 ml of 20% NaCl (w/v) in water for every 20 ml of stain.
Can Coomassie Brilliant Blue interfere in SDS-PAGE?
CBB250 binds proteins and changes their mobility in SDS PAGE.
How do you Destain Coomassie Blue gel?
Destain the gel by soaking for at least 2 hours in 10% acetic acid, 50% methanol, and 40% H2O with at least two changes of this solvent. If the gel still has a Coomassie Blue background then continue destaining until the background is nearly clear.
What is the advantage of using Coomassie Blue over these other methods?
Coomassie dye staining is especially convenient because it involves a single ready-to-use reagent and does not permanently chemically modify the target proteins.
Can you over stain with Coomassie?
If you destain it too long, it will decrease your band intensity and alter your results. Over-destaining happens very often. If it is for quantifying, I would do a 30 minute stain, with a 2-3 hour destain.
Can you reuse Coomassie?
The Coomassie stain is removed by decanting. You may reuse the stain so pour it into a new vial. Cover the gel with the destain solution and allow the gel to destain with gentle agitation. The destain solution should be changed several times, removing it at each change by aspiration.
How do you stain SDS-PAGE with Coomassie blue?
Stain: Dissolve 0.4g of Coomassie blue R350 in 200 mL of 40% (v/v) HPLC grade methanol in water with stirring as needed. Filter the solution to remove any insoluble Page 2 material. Add 200mL of 20% (v/v) acetic acid in water.
Why do we use Coomassie blue in SDS-PAGE?
Coomassie blue dyes are a family of dyes commonly used to stain proteins in SDS-PAGE gels. The gels are soaked in dye, and excess stain is then eluted with a solvent (“destaining”). This treatment allows the visualization of proteins as blue bands on a clear background.
Can you reuse Coomassie stain?
Can you Destain SDS-PAGE gel overnight?
Add Destain, and let destain overnight by gently shaking at Room Temperature.
Does Biobio-Rad offer Coomassie stains?
Bio-Rad offers Coomassie stains in three major formats. Coomassie blue dyes are a family of dyes commonly used to stain proteins in SDS-PAGE gels. The gels are soaked in dye, and excess stain is then eluted with a solvent (“destaining”). This treatment allows the visualization of proteins as blue bands on a clear background.
What is Bio-Safe™ Coomassie?
Environmentally friendly formulation — no addition of methanol or acetic acid required; eliminates the need for hazardous waste disposal Bio-Safe™ Coomassie (161-0786) — Bio-Safe Coomassie Brilliant Blue G-250 stain is fast, simple, sensitive, and convenient.
What is the best stain for Coomassie?
Bio-Safe™ Coomassie (161-0786) — Bio-Safe Coomassie Brilliant Blue G-250 stain is fast, simple, sensitive, and convenient. Coomassie Brilliant Blue R-250 (161-0436) — Coomassie Brilliant Blue R-250 staining solution is the fastest and easiest way to Coomassie-stain Criterion™ precast gels or other polyacrylamide protein gels.
What is Coomassie blue dye used for?
Coomassie blue dyes are a family of dyes commonly used to stain proteins in SDS-PAGE gels. The gels are soaked in dye, and excess stain is then eluted with a solvent (“destaining”). This treatment allows the visualization of proteins as blue bands on a clear background.