Bradford reagent: dissolve 100 mg Coomassie Brilliant Blue G-250 in 50 ml 95% ethanol, add 100 ml 85% (w/v) phosphoric acid. Dilute to 1 liter when the dye has completely dissolved, and filter through Whatman #1 paper just before use.
The Bradford protein assay is a time-tested colorimetric assay. When the Bradford reagent (acidified Coomassie Brilliant Blue G-250) binds to proteins, the dye undergoes a color change in the visible spectrum, with the absorbance maximum moving from 470 to 595 nm. The absorbance at 595 nm is then read either in a spectrophotometer or a microplate reader and is directly proportional to the amount of …
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The Bradford reagent is an acidified solution of Coomassie G-250; the dye is thus primarily protonated and red. The basis for the Bradford assay is that in order for the Coomassie dye to bind stably to protein, it needs to be doubly protonated.
The Bradford Reagent can be used to determine the concentration of proteins in solution.
The Bradford Reagent requires no dilution and is suitable for micro, multiwell plate, and standard (cuvet) assays. The linear concentration range is 0.1-1.4 mg/mL of protein, using BSA (bovine serum albumin) as the standard protein. When using Product B6916, Bradford Reagent, how soon after my color development do I need to read my assay?
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Bradford assay principles Use of Coomassie G-250 Dye in a colorimetric reagent for the detection and quantitation of total protein was first described by Dr. Marion Bradford in 1976. pierce coomassie bradford protein assays are modifications of the reagent first reported by Dr. Bradford. Chemistry of Bradford, Coomassie-based protein assays
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