Coomassie Brilliant Blue Dyes 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 electropho resis and Bradford-type assay reagents for protein quantitation. Typically, coom assie gel stains and protein assay reagents are formulated as very acidic soluti ons in 25 to 50% methanol In acidic conditions, the dye binds to proteins primar ily through basic amino acids (primarily arginine, lysine and histidine), and th e number of coomassie dye ligands bound to each protein molecule is approximatel y proportional to the number of positive charges found on the protein. R-250 (Red-tinted) form lacks two methyl groups G-250 (Green-tinted) form, which is also called colloidal coomassie dye g-250 is more specific in its binding properties and, hence, less sensitive than r-250. g-250 is a greenish-blue when used for staining. it is brown and turns blue (abs orbance wavelength shift) when used for the bradford protein assay. r-250 is dark blue (with a little reddish component) when used to stain. it is n ot used for the bradford assay. g-250 is also the dye of choice for modified coomassie brilliant blue staining m ethods (eg: colloidal, blue silver). Coomassie stain 0.500 g Brilliant Blue 500 mL methanol 100 mL glacial acetic acid 400 mL dH20 Mix well; store at room temperature; can be reused 2-3 times

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