Bradford Assay for Protein
1. Prepare dye reagent by diluting 1 part of Dye Reagent Concentrate with 4 parts of
dd water. Filter to remove particulates. This diluted reagent may be used for up to
one month when kept at room temperature.
2. Add 20l (suggested dilutions) of sample to 1ml of diluted Dye Reagent, vortex
and incubate at room temperature at least 5 minutes. Assay protein solutions in
triplicate.
3. Measure absorbance at 595 nm.
4. Standard Curve:

Prepare 6 dilution of a protein standard -globulin (stock 1 mg/ml)
Amount of the protein [g]
0
4
8
12
16
20

standard [l]
0
4
8
12
16
20
dd water [l]
20
16
12
8
4
0
c [g/l]
0
0.2
0.4
0.6
0.8
1.0
Add 20l of dilution to 1ml of diluted Dye Reagent, vortex and incubate
at room temperature at least 5 minutes. Assay in triplicate.
Reference:
of protein. Anal Chem 72: 248-254