You will prepare 5 cuvettes with different concentration of sodium phosphate (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 0.05mM. Fill the table below (before coming to lab) and use it to prepare your cuvettes. Your blank for each reaction is the PNPP mixed with Tris-HCL and Pi. DO NOT ADD ENZYME AT THIS POINT!!! 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 0.05 100 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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Chemical equilibrium and ionic equilibrium are two major concepts in chemistry. Ionic equilibrium deals with the equilibrium involved in an ionization process while chemical equilibrium deals with the equilibrium during a chemical change. Ionic equilibrium is established between the ions and unionized species in a system. Understanding the concept of ionic equilibrium is very important to answer the questions related to certain chemical reactions in chemistry.
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- You will prepare 5 cuvettes with different concentration of sodium phosphate (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 0.05mM. Fill the table below (before coming to lab) and use it to prepare your cuvettes. Your blank for each reaction is the PNPP mixed with Tris-HCL and Pi. DO NOT ADD ENZYME AT THIS POINT!!!
|
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 |
0.05 |
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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