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Bradford Assay Lab Report

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Bradford Assay Lab Report
During this experiment, the method of Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) was conducted using a negatively charged protein and pre-stained molecular weight markers. The hypothesis was that the molecular weight of N-acetyl-β-D-hexosaminidase B would be 28,000 kDa. To confirm or reject the hypothesis, the molecular weight of N-acetyl-β-D-hexosaminidase B and the concentration of protein had to be determined. The electrophoresis of the protein gel were conducted using a Hoefer SE 600 Ruby at 30 mA and stained with Coomassie Brilliant Blue Dye for 4 hours. SDS-PAGE Slab Gel Destaining Solution was then used for the removal of excess dye in the protein gel for 16 hours. In order to determine the Rf values of the protein bands, the protein gel had to be scanned using a Ricoh Super G3 scanner. Calculating the Rf values and log molecular weight values produced a standard curve with the equation of y=-1.1067x+5.2587. The antilog of the log molecular weight calculated gave a molecular weight of 79,432 kDa.
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