Comparative analysis of different methods for the determination of unknown protein concentration

Protein is a class of nitrogenous organic compounds that consist of large molecules composed of one or more long chains of amino acids and are an essential part of all living organisms, especially as structural components of body tissues such as muscle, hair, collagen, etc., and as enzymes and antibodies. It maintains the cell shape, catalyze the metabolic reactions and properties, and regulate gene expression with the help of its functional group and also essential in animal diets. A rapid and accurate method for the estimation of protein concentration is essential in many fields of protein study. There are different methods for protein quantization they are Lowry methods, Bradford method, Biuret assay method, Direct absorbance method etc. In this method we prepare different concentration protein solution and also take unknown protein solution and measure there absorbance at a fixed wavelength. From the spectrum we get the unknown protein solutions absorbance. Then we plot a graph where we plot absorbance vs. concentration (mg/mL) at a fixed wavelength and we get a calibration curve, from the graph we can determine the molar extinction co-efficient. Finally we establish a new method arbitrarily named as Crocoite method for protein quantization which follows the same process and in this method we measure the absorbance at 632nm. Finally, we compare Biuret assay method, direct absorbance method and Crocoite method by molar extinction co- etficient value . And we see that Crocoite method is moresensible than other two methods. So, we can quantatize protein more accurately by the Crocoite method.

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