- Prepare a standard curve of five to six concentrations of a protein standard, which is representative of the protein solution to be tested. Usually 0 – 10 ug of protein adjusted to 800 uL with water or sample buffer.
- Pipet 10 uL each sample into a tube and adjust volume to 800 uL (NOTE: the volume of the unknown can be adjusted to as much as 800 uL). Samples are usually assayed in duplicate or triplicate.
- Add 200 uL of the dye reagent (DO NOT DILUTE). Vortex each tube.
- Incubate at room temperatre for at least 5 minutes. Absorbance will increase over time; samples should incubate at room temperature for no more than 1 hour.
- Measure absorbance at 595 nm