Expt3 Bradford Assay- Sample Journal Article Report F09

It was seen that the bradford assay is a quick and

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Unformatted text preview: 80 μg/mL. It was seen that the Bradford assay is a quick and effective method for determining an unknown protein concentration, and that this method is highly sensitive, able to be effective around concentrations of about 1 μg/mL. Introduction: The Bradford assay is a very popular protein assay method because it is simple, rapid, inexpensive and sensitive. The Bradford Assay is based on protein binding of Coomassie Brilliant Blue G-250 dye to protein residues in acidic solution and provides various advantages over other methods. The dye is believed to bind especially strongly to arginine residues and less strongly to histidine, lysine, tyrosine, tryptophan, and phenylalanine residues where the binding of dye to protein in acidic solution causes a shift in wavelength of maximum absorption of the dye from 465 nm to 595 nm that is linear to protein concentration.1, 2 The absorption at 595 nm, then, is directly related to the concentration of the protein. 1, 2 Generally, a calibration curve is prepared using bovine plasma gamma globulin or bovine serum albumin as a standard. The assay requires only a single reagent, an acidic solution of Coomassie Brilliant Blue G-250 shown below:4 After addition of dye solution to a protein sample, color development is complete in 2 minutes, and the color remains stable for up to 1 hour. With a micro assay procedure, the Bradford assay can be used to determine proteins in the range of 1...
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