How do you make colloidal Coomassie?
first dissolve aluminum sulfate in Milli-Q water. thereafter add ethanol, homogenize, and mix CBB G-250 to the solution. as recently as the solution is completely dissolved, add phosphoric acid (the incorporation of the acid to the alcoholic media lets the Coomassie molecules aggregate into their colloidal state)
How do you make Coomassie blue destaining solution?
Prepare a destain solution containing 10% ethanol and 7.5% acetic acid. Place one or two stained gels in a staining container containing the 100 ml destain solution. Loosely cover the staining container and heat in a microwave oven at full power for 1 minute.
What is Coomassie staining for?
Description. 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 Coomassie stain overnight?
Coomassie blue staining of proteins Submerge the gel in the Commassie blue stain solution. Use just enough to completely submerge the polyacrylamide gel. Stain for 1-4 hours or overnight at room temperature with gentle shake. Coomassie blue stain solution can be reused for serveral times.
What is the difference between R250 and G250 Coomassie Brilliant Blue G-250?
The more popular is Coomassie R250 (Reddish tinted blue) for electrophoresis (more sensitive: can detect as little as 0.1 µg of protein), and Coomassie G250 (Greenish tinted blue) for protein assay in solutions (because it is more convenient – soluble).
Which amino acids does Coomassie bind to?
In acidic conditions, Coomassie dye primarily binds basic amino acids (arginine, lysine and histidine).
What does Coomassie blue bind to?
Is Coomassie Blue toxic?
Coomassie Briliant Blue R-250 Dye (6104-59-2) Potential Adverse human health effects and symptoms : Non-toxic if swallowed (LD50 oral, rat > 5000 mg/kg). Not irritant to skin. Not irritant to eyes.
Why do we stain with Coomassie blue?
Coomassie Blue stain is used to stain the protein bands in polyacrylamide gels. The dye binds more tightly to the proteins than the to the gel matrix, however, so the dye can subsequently be removed from only the protein-free parts of the gel using a similar solvent from which the dye is omitted. This is the destain.
What is the function of Coomassie blue?
Why do we use Coomassie blue?
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 Coomassie blue go down the drain?
COOMASSIE BLUE destain may contain alcohol* and cannot be disposed of in the sink! Hazardous Materials are NEVER to be disposed of in the sink!
What is the recipe for colloidal Coomassie blue stain?
Colloidal Coomassie Blue Stain: Recipe and Protocol I learned this Colloidal Coomassie Blue staining recipe and protocol when I was a postdoc at Harvard Medical School.. Rumor has it, a postdoc reverse engineered a “safe stain”…
What is the absorption rate of Coomassie blue?
At a pH of around 1 the dye bears an overall charge of +1 (the 2 sulfonic acid groups are negatively charge, thanks to theirextremely low pKa), so the dye is green with an absorption maximum at 620 nm while above pH 2 the dye is bright blue with a maximum at 595 nm.
Which is more sensitive, Coomassie blue or G250?
Coomassie Blue is widely used in visualizing proteins separated by either agarose or acrylamide gel electrophoresis. The more popular is Coomassie R250 (Reddish tinted blue) for electrophoresis (more sensitive: can detect as little as 0.1 µg of protein), and Coomassie G250 (Greenish tinted blue) for protein assay in solutions (because it is more