The concentration of purified OXA-M290 is tested with a BCA assay. Serial dilutions of a bovine serum albumin (BSA) stock solution are prepared, then pipetted into a 96-well plate; each dilution of the BSA standard is tested in triplicate. Then, bicinchoninic acid and Cu2+ ions are added to all of the wells of the plate. After incubating the plate for 1 hour, a microplate reader is used to measure the absorbance of all of the wells in the plate at 560 nm. This generates the following data: BSA conc. (μg/mL), Replicate 1 Absorbance, Replicate 2 Absorbance, Replicate 3 Absorbance 40, 1.360, 1.403, 1.481 20, 0.750, 0.745, 0.810 10, 0.380, 0.344, 0.398 5, 0.198, 0.160, 0.183 2.5, 0.090, 0.100, 0.085 1.25, 0.038, 0.043, 0.051 0.625, 0.024, 0.028, 0.019 Prepare a calibration curve using these data. You can use Excel, R, SPSS or an equivalent graphing software. In this graph, plot absorbance (y-axis) against the concentration of the protein standard (x-axis). Calculate and plot the average absorbance values for the three replicates that were measured for each BSA concentration; you do not need to plot each replicate separately. Your graph should include a trendline, an equation for the trendline, and an R2 value. Your graph should also include a title at the top, and appropriate titles for the x- and y-axes. Be sure to provide the equation for the trendline for your graph and the image of your graph.
The concentration of purified OXA-M290 is tested with a BCA assay. Serial dilutions of a bovine serum albumin (BSA) stock solution are prepared, then pipetted into a 96-well plate; each dilution of the BSA standard is tested in triplicate. Then, bicinchoninic acid and Cu2+ ions are added to all of the wells of the plate. After incubating the plate for 1 hour, a microplate reader is used to measure the absorbance of all of the wells in the plate at 560 nm. This generates the following data:
BSA conc. (μg/mL), Replicate 1 Absorbance, Replicate 2 Absorbance, Replicate 3 Absorbance
40, 1.360, 1.403, 1.481
20, 0.750, 0.745, 0.810
10, 0.380, 0.344, 0.398
5, 0.198, 0.160, 0.183
2.5, 0.090, 0.100, 0.085
1.25, 0.038, 0.043, 0.051
0.625, 0.024, 0.028, 0.019
Prepare a calibration curve using these data. You can use Excel, R, SPSS or an equivalent graphing software.
In this graph, plot absorbance (y-axis) against the concentration of the protein standard (x-axis). Calculate and plot the average absorbance values for the three replicates that were measured for each BSA concentration; you do not need to plot each replicate separately. Your graph should include a trendline, an equation for the trendline, and an R2 value. Your graph should also include a title at the top, and appropriate titles for the x- and y-axes.
Be sure to provide the equation for the trendline for your graph and the image of your graph.
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