Bovine Hydrochloric Acid Solution

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During this experiment that was conducted in partners, the concentration of RNase H that was purified in the previous experiment #6 was determined and also the concentration of an unknown given solution using their absorbance values obtained from the spectroscopy apparatus. These concentrations were calculated using the equation of the line produce with the values of the concentration (x-axis) and corresponding absorbance (y-axis) of five protein bovine gamma globulin standard solutions. In order to prepare each of these standards within a range of 50 microliters to 1200 microliters and using a provided stock solution of 1.46 mg/mL of the protein bovine gamma globulin (IgG) different calculations were performed in order to find out the exact …show more content…
Bradford for protein determination was used in the lab in order to determine the concentration of the unknown 2 and the previously purified protein. This protein dye complex is the reason for a shift in the dye absorption with maximum between 465 to 596 nm. This derived amount of absorption does not require dilution and can be used for both microassays and standard solutions. The analysis provides a linear response with concentrations of Bovine Serum Albumin (BSA) from 1 to 140 microliters. The reagent used was the combination of Bradford Reagent-Brilliant Blue G in phosphoric acid and methanol. The dye molecule is mostly deprotonated and converted on the process to a reddish colored species that have the potential to bind to other proteins. Once this red colored species bind to a protein the bound dye molecule transforms to blue species, and this color change is the one measure by the spectrophotometrical. The assay is a spectroscopy analytical technique based upon the absorbance maximum for an acidic solution of Coomassie Brilliant Blue. From the assay is observed the maximum absorbance for an acidic solution of Coomassie Brilliant Blue G-250 mostly shifts from 465 to 595 nm upon binding to protein studied occurs. Both ionic interaction and hydrophobic are used to stabilize the anionic form of the dye, which experience a visible change in the color (Stoscheck). This assay is performed at room temperature and a change in color begins immediately. It has great utility due to the fact that the extinction coefficient of a dye-protein complex solution remains constant for a range of over 10-fold concentration

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