Protein Assay

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Protein Assay by(Name): Course: Tutor: Institution: City/State: Date: Protein Assay Purpose The purpose of this lab exercise was to determine the protein concentration of solutions by the Lowry and Bradford Assays. Materials and Methods/Procedure Materials Materials include a spectrophotometer, vortex mixer, cuvettes, micropipettes, marker pen, water baskets, distilled water and micropipette tips. They also include Bovine Serum Albumin (BSA) standard solution, Sodium Phosphate buffer, Lowry reagents A and B, Bio-Rad Dye reagent solution and BSA solution of unknown concentrations. Methods Part I. Lowry Assay. A series of 6 Eppendorf tubes containing a wide range of the protein standard solution was prepared to a final volume of 25uL and labeled. A control tube containing 25uL of water was also added. A series of tubes ranging from 0 to 5 to 40ug BSA were made. This was followed by preparation of a series of dilutions with unknown concentration by adding water in different volumes to a final 25uL. A 125uL Reagent A was added to each microfuge tube and mixed thoroughly with a vortex, followed by the addition of 1000uL of Reagent B to each tube and an immediate and thorough vortex. The solution was incubated for 15 minutes and absorbance’s read at 650nm after blanking the spectrophotometer with tube #1. Part II.A 760uL solution of 0.01mg/L BSA standard was prepared by diluting a 2.0mg/L of the standard solution with 10mM NaH2PO4 AT pH 7.0. This was followed by the preparation of standard serial solutions with a final volume of 800uL. A blank tube containing 800uL of water was used as control. A set of six tubes with varying dilutions was then

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