Bradford Method: Colorimetric Protein Assay

Bradford method is a common colorimetric method to determine protein concentration in a sample solution. The Bradford method of protein determination is based on the binding of a dye, Coomasie Blue G, to the protein. This binding shifts the absorbtion maximum of the dye from red to blue. The absorbance of the solution is measured at 595 nm and is proportional to protein concentration when compared to a standard curve. Two types of assay are described here: the standard assay, which is suitable for measuring between 10 and 100 microgram of protein, and the microassay, which detects between 1 and 10 microgram of protein.

Quantitative Estimation of DNA Concentrations

DNA, RNA, and protein strongly absorb ultraviolet light in the 260 to 280 nm range. UV spectroscopy can be used as a quantitative technique to measure nucleic acid concentration and protein contamination. Nucleic acids strongly absorb at 260 nm and less strongly at 280 nm while proteins do the opposite. The general rules for determining the concentrations of nucleic acids at 260 nm are:

Estimation of DNA Concentrations

The determination of the concentration of DNA or RNA in solution is a fundamental task in molecular biology. DNA is usually the limiting reagent in most experiments; therefore, the knowledge of its concentration is critical. Determination of the DNA concentration can be estimated either by qualitatively comparing the fluorescence of DNA bands in an agarose gel to a standard or by spectrophotometric means.

Measurement of Biosurfactant Activity

Biosurfactant activity was measured by an oil displacement test. This method is so sensitive that only a small amount of sample is required to measure the surfactant activity. Here is the method:

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