8
CH 461 & CH 461H F ‘06
October 18, 2006 EXP306.wpd Ingle, Pastorek, Westall
II. SOLUTIONS
A. Stock Solutions
The following stock solutions are available:
Buffer Solution (to control the pH of the reaction mixture)
42 7 2 3 4
• 0.024 M pyrophosphate pH 8.8 buffer (made from Na P O C10H O and H PO )
ADH Solution (the enzyme solution)
•X :g/mL ADH in a 0.01 M pH 7.5 potassium phosphate buffer (you will be provided the
value of X which is typically 1 to 15).
• Dilute solutions of ADH rapidly lose activity upon standing and are quite unstable at pH
values below 6.0 or above 8.5. The deactivation rate is reduced by storing the solutions at
0-5 C when not in use and by placing the enzyme solution container in an ice bath during
lab work.
NAD Solution (coenzyme)
+
2
• 0.017 g/mL (2.5 mM) in H O
• This solution is stored in the refrigerator when not in use and in an ice bath during lab
work.
Ethanol Solutions
• 0.2500 M (14.58 mL of 100% ethanol (MW, 46.07; density, 0.790) / L)
• 0.1000 M
• 0.01250 M
• 0.003125 M
• These stock solutions of ethanol are prepared in 0.024 M, pH 8.8 pyrophosphate buffer.
