Protein Determinations

Protein determination by the Bradford method

 


Bradford protein assay

Considerations for use

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.

We use a kit Cat.#23200 from Pierce Chemical Co. to determine protein concentrations in rhesus monkey follicular fluid.

Principle

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.

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Last updated: Thursday, December 7, 2000

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