How do I prepare a solution of Coomassie R-250 dye?

How do I prepare a solution of Coomassie R-250 dye?

How do I prepare a solution of Coomassie R-250 dye? Add 100 mL of glacial acetic acid to 450 mL. Dissolve 3 g of Coomassie R-250 dye in 450 mL methanol. Mix the acetic acid and methanol solutions.

How do you make Coomassie destaining solution?

Mix H2O, methanol, and acetic acid in a ratio of 50/40/10 (v/v/v).

How do you dilute a Coomassie?

Dissolve 100 mg Coomassie Brilliant Blue G-250 in 50 ml of 95% ethanol and add 100 ml of 85% phosphoric acid while stirring continuously. When the dye has dissolved dilute to 1 l in water.

How to make Coomassie Brilliant Blue staining solution?

Recipe. Coomassie Brilliant Blue staining solution. Dissolve 1 g of Coomassie Brilliant Blue (Bio-Rad) in 1 liter of the following solution: Methanol (50% [v/v]) Glacial acetic acid (10% [v/v]) H 2 O (40%) Stir the solution for 3-4 hours and then filter through Whatman filter paper. Store at room temperature.

How long to stain gel with Coomassie Blue R-250?

Stain gel in 10% Acetic Acid in water, containing 60 mg/L of Coomassie Blue R-250. Bands will appear in 30 minutes. Allow staining to proceed until desired band intensity is reached. In this protocol, background staining is low due to the very low dye concentration used.

Which is the first dye in the Coomassie series?

The first of the Coomassie series was Coommassie Blue R-250 (“R” standing for “reddish” and “250” being the dye strength indicator). Coommassie Blue G-250 (“G” for “greenish”) and Coomassie Violet R-150 later followed. The most commonly used dyes in the laboratory for staining PAGE gels are Coomassie Blue R-250 and G-250.

How does Coomassie Brilliant Blue G-250 bind to proteins?

When Coomassie Brilliant Blue G-250 binds to proteins in acid solution, it has an absorbance shift from 465 nm to 595 nm. The absorbance data can then be used in Beer’s law to determine protein concentration and ultimately the actual amount of protein in a given solution.