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1. You will use 0.5mM PNPP, 0.2M Tris-HCl, and 1mM P; stock solutions to prepare two sets of five samples (Sets A and B), each with varying P; concentrations. The PNPP concentration for all samples in Set A will be 0.05mM; and 0.150mM for all samples in Set B. Fill out the tables below and use them to prepare your samples. Be aware of the following: • Prepare the samples directly in cuvettes. Prepare one sample at a time to minimize exposing PNPP to air. You can prepare the next sample while the current one is being measured. • Each sample before adding enzyme (i.e., the cuvette only contains PNPP, Pi, and Tris-HCI) is its own blank. In other words, you add the appropriate amounts of PNPP, Pi, and Tris- HCI to a cuvette, press "Measure Blank", then add 100μL of AP to the same cuvette, mix, and press "Measure Sample". • You must blank the instrument (before adding the enzyme) each time you prepare a new sample. Table 1. Sample Set A (PNPP concentration = 0.05mM) Final [Pi] (μM) Vol of 1mM Pi (μL) [PNPP] (mM) Vol of 0.5mM PNPP (μL) Vol of 0.2M Tris-HCI (μL) Vol of enzyme Total Vol (μL) (μL) 0 0.05 100 1500 3.33 0.05 100 1500 20 0.05 100 1500 50 0.05 100 1500 100 0.05 100 1500

1. You will use 0.5mM PNPP, 0.2M Tris-HCl, and 1mM P; stock solutions to prepare two sets of five samples (Sets A and B), each with varying P; concentrations. The PNPP concentration for all samples in Set A will be 0.05mM; and 0.150mM for all samples in Set B. Fill out the tables below and use them to prepare your samples. Be aware of the following: • Prepare the samples directly in cuvettes. Prepare one sample at a time to minimize exposing PNPP to air. You can prepare the next sample while the current one is being measured. • Each sample before adding enzyme (i.e., the cuvette only contains PNPP, Pi, and Tris-HCI) is its own blank. In other words, you add the appropriate amounts of PNPP, Pi, and Tris- HCI to a cuvette, press "Measure Blank", then add 100μL of AP to the same cuvette, mix, and press "Measure Sample". • You must blank the instrument (before adding the enzyme) each time you prepare a new sample. Table 1. Sample Set A (PNPP concentration = 0.05mM) Final [Pi] (μM) Vol of 1mM Pi (μL) [PNPP] (mM) Vol of 0.5mM PNPP (μL) Vol of 0.2M Tris-HCI (μL) Vol of enzyme Total Vol (μL) (μL) 0 0.05 100 1500 3.33 0.05 100 1500 20 0.05 100 1500 50 0.05 100 1500 100 0.05 100 1500

Chemistry
Chemistry
10th Edition
ISBN:9781305957404
Author:Steven S. Zumdahl, Susan A. Zumdahl, Donald J. DeCoste
Publisher:Steven S. Zumdahl, Susan A. Zumdahl, Donald J. DeCoste
Chapter1: Chemical Foundations
Section: Chapter Questions
Problem 1RQ: Define and explain the differences between the following terms. a. law and theory b. theory and...
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1. You will use 0.5mM PNPP, 0.2M Tris-HCl, and 1mM P; stock solutions to prepare two sets of five samples (Sets A and B), each with varying P; concentrations. The PNPP concentration for all samples in Set A will be 0.05mM; and 0.150mM for all samples in Set B. Fill out the tables below and use them to prepare your samples. Be aware of the following: • Prepare the samples directly in cuvettes. Prepare one sample at a time to minimize exposing PNPP to air. You can prepare the next sample while the current one is being measured. • Each sample before adding enzyme (i.e., the cuvette only contains PNPP, Pi, and Tris-HCI) is its own blank. In other words, you add the appropriate amounts of PNPP, Pi, and Tris- HCI to a cuvette, press "Measure Blank", then add 100μL of AP to the same cuvette, mix, and press "Measure Sample". • You must blank the instrument (before adding the enzyme) each time you prepare a new sample. Table 1. Sample Set A (PNPP concentration = 0.05mM) Final [Pi] (μM) Vol of 1mM Pi (μL) [PNPP] (mM) Vol of 0.5mM PNPP (μL) Vol of 0.2M Tris-HCI (μL) Vol of enzyme Total Vol (μL) (μL) 0 0.05 100 1500 3.33 0.05 100 1500 20 0.05 100 1500 50 0.05 100 1500 100 0.05 100 1500
Transcribed Image Text:1. You will use 0.5mM PNPP, 0.2M Tris-HCl, and 1mM P; stock solutions to prepare two sets of five samples (Sets A and B), each with varying P; concentrations. The PNPP concentration for all samples in Set A will be 0.05mM; and 0.150mM for all samples in Set B. Fill out the tables below and use them to prepare your samples. Be aware of the following: • Prepare the samples directly in cuvettes. Prepare one sample at a time to minimize exposing PNPP to air. You can prepare the next sample while the current one is being measured. • Each sample before adding enzyme (i.e., the cuvette only contains PNPP, Pi, and Tris-HCI) is its own blank. In other words, you add the appropriate amounts of PNPP, Pi, and Tris- HCI to a cuvette, press "Measure Blank", then add 100μL of AP to the same cuvette, mix, and press "Measure Sample". • You must blank the instrument (before adding the enzyme) each time you prepare a new sample. Table 1. Sample Set A (PNPP concentration = 0.05mM) Final [Pi] (μM) Vol of 1mM Pi (μL) [PNPP] (mM) Vol of 0.5mM PNPP (μL) Vol of 0.2M Tris-HCI (μL) Vol of enzyme Total Vol (μL) (μL) 0 0.05 100 1500 3.33 0.05 100 1500 20 0.05 100 1500 50 0.05 100 1500 100 0.05 100 1500
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