* Coomassie Brilliant Blue G-250 Dye: The assay uses this dye, which binds to proteins. This binding changes the dye's structure and causes a shift in its absorbance spectrum.
* Color Change: When the dye binds to protein, it changes color from brown to blue. This blue form has a maximum absorbance at 595 nm.
* Quantifying Protein: The intensity of the blue color is directly proportional to the amount of protein present. By measuring the absorbance at 595 nm, we can quantify the protein concentration.
In summary:
* The Bio-Rad assay uses Coomassie Brilliant Blue G-250 dye to bind to proteins.
* The bound dye turns blue and absorbs maximally at 595 nm.
* Absorbance at 595 nm is measured to determine the concentration of protein.
Therefore, while the absorbance is measured at 595 nm, it's not the total protein being measured directly, but the color change caused by the dye binding to proteins.
