User:Dhea Patel/Notebook/CHEM 572: ADA&Inhibitor Kinetics/2013/01/23

Objective

• To complete project proposal
• To complete experimental protocol for next week.

Experimental Protocol

Protocol for 0.1 M Sodium Phosphate Buffer (pH 7.4)

• Add 3.1 g of NaH2PO4•H2O and 10.9 g of Na2HPO4 (anhydrous) to distilled H2O to make a volume of 1 L.
  • The pH of the final solution will be 7.4.
  • This buffer can be stored for up to 1 mo at 4°C.
• Dilute to 0.05M: (take 50 mL of 1M solution and dilute in 1 Liter H2O)

Running ADA Kinetics

• The following table outlines the concentrations and volumes of the solutions used in the ADA kinetics assay to be performed next Tuesday.

• To prepare a 30 mM stock solution of adenosine:

of adenosine × = of adenosine in buffer

• To make 10mL of solution, 0.080172g of adenosine will be added to 10mL of phosphate buffer.