Bradford Assay Essay

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Quiz 9 Question Pool Below please find the ninth of the weekly lab quizzes. All 10 points of the quiz will come from this list of questions. 1. Describe in detail the assay you will do this week on the cellular fractions from last week. (5 points) 2. Fill in the following table so that all tubes have a final volume of 60 μl: (5 points) Assay Tube # Desired [BSA](mg/ml) Volume of 1 mg/mlBSA (μl) Volume of phosphate buffer (μl) 1 = blank 0 0 60 2 0.2 3 0.4 4 0.6 5 0.8 6 1.0 3. What does the Bradford assay measure? (1 point) 4. Which dye does the Bradford assay use? (1 point) 5. To determine the total protein content in a particular fraction, what must you multiply by what? Hint: there are 3 quantities!…show more content…
In the slides, the intensity of staining of _______________________________ is important. (1 point) 7. What is enzyme activity? (2 points) 8. What is the significance of total activity? (3 points) 9. What is the significance of specific activity? (3 points) 10. In what units will we measure enzyme activity? (2 points) 11. Which 2 enzymes are we following & where is each found? (4 points) 12. a. With which instrument will we measure the amount of product formed by the enzymatic reactions? (1 point) b. What property must a substance have in order to be measured by the instrument in part (a)? (1 point) c. What is the measurable product of each enzyme reaction? (2 points) 13. For an enzymatic reaction, what should a blank contain? Hint: the answer is NOT “extra phosphate buffer”! (2 points) 14. What substances will you add to each enzyme reaction to stop it? What do these substances do? (5 points) 15. a. For succinate dehydrogenase, what volumes of enzyme go into the reactions? There are 2 different amounts, corresponding to 2 different dilutions. (2 points) b. If after 30 mins you measure an absorbance of 0.65 for a particular fraction at the lesser dilution (greater amount of fraction used), what is the activity*/ml of that fraction? Show your work! (2

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