The Bradford assay is very fast and uses about the
same amount of protein as the Lowry assay. It is fairly accurate and
samples that are out of range can be retested within minutes. The
Bradford is recommended for general use, especially for determining
protein content of cell fractions and assesing protein concentrations
for gel electrophoresis.
The assay is based on the observation that the absorbance
maximum for an acidic solution of Coomassie Brilliant Blue G-250 shifts
from 465 nm to 595 nm when binding to protein occurs. Both hydrophobic
and ionic interactions stabilize the anionic form of the dye, causing
a visible color change. The assay is useful since the extinction coefficient
of a dye-albumin complex solution is constant over a 10-fold concentration
range.