Bradford Assay Protocol

Our Bradford assay reagents are found in the Quick Start Bradford Protein Assay Kit 2 from BioRad, part number 500‐0202. The kit contains the BSA standard set and the 1x dye reagent.

When performing a Bradford assay, use a UV Flat Bottom Microtiter Plate from Thermo, part number 8404. The microtiter plates are in a 96‐well format.

For the calibration curve, pipet 5 uL in duplicate for each standard in the BSA standard set. Each 5 uL sample goes in a separate well. There are seven standards in the set, which are:

0.125 ug/uL
0.25 ug/uL
0.5 ug/uL
0.75 ug/uL
1 ug/uL
1.5 ug/uL
2 ug/uL

When pipetting out the calibration curve, make sure to include two blank wells for the zero point in the calibration curve.

Add 5 uL of each sample to a separate well. If you have enough sample, adding 5 uL in duplicate is preferable.

Add 245 uL of the 1x dye reagent to every well that contains a sample. Remember to add dye reagent to the blank (zero point) wells.

Read the measurements on a plate reader set to a wavelength of 595 nm.