Burden, Sierra, BioChem, Friday G, PostLab

Burden, Sierra, BioChem, Friday G, PostLab - Student...

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Student Name: ________Sierra Burden_______________ CA & Intern Name: _______Tripp________________ Section Day and Letter: _________Friday G______________ E-mail Address: [email protected]_______________ Post-Lab Worksheet (20 points) This post-lab write-up is intended to guide the interpretation of your data as well and concludes concepts covered in our lab. Data Analysis 1. Based on your standard curves of the Bradford and ONPG assays performed in week 1, what were your total and β-galactosidase protein concentration? What information would you need to be able to calculate a percent purity? Can you do this with our lab? Paste your standard curves into the space below and explain your answers. (2 pts) The total protein concentration was .43 mg/mL and the B-galactosidase concentration was .42 units/ 40 ul. The percent purity can be calculated by determining the concentration of a pure B-galactosidase sample. The concentration of the pure sample would be divided by the concentration of the experimental sample. 2. What conditions did you test for your storage conditions (i.e., what was temperature 1, temperature 2)? At which temperature did the enzyme fare better? Did the addition of the BME and Mg 2+ appear to make a difference in the activity of your enzyme? Explain your answer. (1 pt) The storage conditions tested temperatures 37°C and 4°C. The two different temperatures were also tested with and without BME plus Mg 2+ . At 4°C with BME plus Mg 2+ , more ONP- was produced and the enzyme was more effective. The samples with the BME plus Mg 2+ produced more ONP- than the samples without it. For example, the BME and Mg 2+ at 4°C had an optical reading of .869 versus the sample at the same temperature with an optical reading of .133. The BME plus Mg 2+ , therefore, made the enzyme more effective. 3. Determine your net absorbance values and convert the absorbance to concentrations of ONP. Plot the [ONP-] production over time for all six concentrations of [ONPG] in each enzyme-inhibitor pairing. (You will end up
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