User:Mary Mendoza/Notebook/CHEM572 Exp. Biological Chemistry II/2013/01/29

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Preparation of ADA Kinetic Assay Reactants

  • Adenosine deaminase was ordered from Worthington (19.8 u/mg). To obtain 0.5 M per mL, the calculation below was made amounting to 0.0253.


\frac{0.5 units}{19.8 u/mg} = 0.0253 mg of ADA in 1 mL phosphate buffer


  • For convenience, it was decided to dissolve 2.5 mg into 100 mL of phosphate buffer. The final amount weighed 2.0 mg which will be dissolved into 80 mL of phosphate buffer. ADA was refrigerated between 2-8°C temporarily.
  • In preparation of the sodium phosphate buffer, 3.1093 g of sodium phosphate monobasic (chempure, FW 268.07 g/mol) and 10.9019 g of sodium phosphate dibasic (fisher scientific, FW 137.99 g/mol) were dissolved in 1 L water.
  • The pH of the buffer was adjusted to 7.4 by the addition of 1 M sodium hydroxide.




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