Determination of Protein Assay Using Bradford’s Reagent

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Determination of Protein Assay Using Bradford’s Reagent Priscilla Mariel M. Cadiz *E-mail: cadizpriscillamariel@yahoo.com Different experiments in biology and biochemistry need to know the amount of protein present in a solution also known as protein quantitation. There are 4 types of protein quantitation assay namely Biuret, BCA, Modified Lowry and Bradford Assay. For this experiment, Bradford Assay will be taken into consideration. Bradford Assay is based on a protein dye binding involving Coomassie Brilliant Blue dye, which binds to aromatic amino acids including arginine. Six samples were prepared for this experiment and each sample contains 250ul Bradford Reagent, varying concentration of phosphate buffer saline and different concentrations of albumin. Each sample was then transferred into a cuvette then placed in a spectrophotometer that is set to 595nm which measures the absorbance of each sample and comparing it to its’ concentration. A standard curve was also produced. Results obtained however were not consistent to the Beer-Lambert’s law that indicates that there is a linear or direct relationship between the absorbance and the concentration, which means that as the concentration increases, the absorbance also, increase. Allowing the graph to consistently increase. The values obtained did not consistently increase however its’ Pearson’s Correlation Coefficient is 0.0189 which means that there is still a weak direct relationship between the absorbance and the concentration even if the values are not constantly increasing. Different factors may affect the inconsistency of the data such as failure to follow the protocol consistently. 1. INTRODUCTION Protein quantitation is a procedure in order to determine the amount of protein present in a solution. There are four types of protein quantitation assays namely BCA, Biuret,

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