User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/11/13

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ADA Kinetic Assay Preparations

  • A weight of 0.6702 g of sodium phosphate dibasic was dissolved in 50 mL of water to obtain a molarity of 0.05 M. The pH of the solution was adjusted to pH 7.4.


.050 L of water × [math]\displaystyle{ \frac{0.05 mol}{1L} }[/math] = .0025 mol of Na2HPO4 × [math]\displaystyle{ \frac{268.07 g}{1 mol} }[/math] = 0.6702 g


  • The pH was was adjusted to 7.4 by the addition of 2 drops of 12 M HCl.
  • To obtain 1 mM inosine, 1.5 mg of the solid was dissolved in 1 mL buffer. This was further diluted by collecting 89.3 µL of the 1.5 mg/ml inosine into 5 mL of the sodium phosphate buffer.


.0015 g of inosine × [math]\displaystyle{ \frac{1 mol}{268.2 g} }[/math] = .000005596 mol ÷ .001 L = .005596 M = 5.596 mM


5.596 mM (V1)= 0.1 mM (5 mL)

V1 = 0.08934 mL = 89.3 μL in 5 mL of buffer

  • 3 mM adenosine was prepared by dissolving 0.0082 g of the solid into 10 mL sodium phosphate buffer. The stock concentration of adenosine was 3.07 mM.


.0082 g of adenosine × [math]\displaystyle{ \frac{1 mol}{267.24 g} }[/math] = 3.06840 × 10-5 ÷ .010 L = .00307 M = 3.07 mM



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