What dye-binding method uses an increase of absorbance at 595 nm to determine total protein concentration?

The method that uses an increase of absorbance at 595 nm to determine total protein concentration is the Coomassie Brilliant Blue 250 method. This dye-binding assay relies on the interaction between Coomassie Brilliant Blue dye and proteins, resulting in a colorimetric change that can be quantitatively measured.

When proteins are present, they bind to the dye, causing a shift in the absorption maximum of the dye from 465 nm (when unbound) to 595 nm (when bound to protein). The degree of color change is directly proportional to the protein concentration in the sample. By measuring the absorbance at 595 nm, one can determine the total protein concentration against a standard curve created using known protein concentrations.

This method is highly sensitive and commonly used in laboratories to quantify protein concentration efficiently.