solution (in water) and 0.2M Tris-HCl, pH=8.00. For this part, the concentration of PNPP will remain constant at 0.150mM. Fill the table below (before coming to lab) and use it to prepare your cuvettes. The completed table must be in your pre-lab. Your blank for each reaction is the PNPP mixed with Tris-HCL and Pi. Again, prepare one cuvette at the time to minimize exposure of the PNPP to the air. You can prepare your

Principles of Modern Chemistry
8th Edition
ISBN:9781305079113
Author:David W. Oxtoby, H. Pat Gillis, Laurie J. Butler
Publisher:David W. Oxtoby, H. Pat Gillis, Laurie J. Butler
Chapter15: Acid–base Equilibria
Section: Chapter Questions
Problem 95AP
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  • Prepare another 5 cuvettes with different concentration of Pi using the stock 1mM Pi, 0.5mM PNPP solution (in water) and 0.2M Tris-HCl, pH=8.00. For this part, the concentration of PNPP will remain constant at 0.150mM. Fill the table below (before coming to lab) and use it to prepare your cuvettes. The completed table must be in your pre-lab. Your blank for each reaction is the PNPP mixed with Tris-HCL and Pi. Again, prepare one cuvette at the time to minimize exposure of the PNPP to the air. You can prepare your next cuvette while your previous one is being read. 
  • previous one is being read. 

Final [Pi]uM

Volume of 1mM Pi

Final [PNPP] mM

Volume of 0.5mM PNPP

Volume of 0.2M Tris-HCl (uL)

Volume of enzyme (uL)

Total Volume (uL)

0

 

0.150

   

100

1500

3.33

 

0.150

   

100

1500

20

 

0.150

   

100

1500

50

 

0.150

   

100

1500

100

 

0.150

   

100

1500

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