CRCJ 1000 Lecture 3: DETAILED PDF LEC NOTES - CARLETON UNI - JEFFREY MONAGHAN
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Related Questions
Calculate the volumes of needed to make solutions of alkaline phosphatase and p-nitrophenylphosphate with the given concentration and update the table below
Reagents:
Reaction buffer: 0.5 M Tris HCl (pH 8.0) with 5 mM MgCl2
Alkaline phosphatase: 500 µg/mL stock solution in reaction buffer.
p-nitrophenylphosphate: 1000 µM and 10,000 µM stock solution in reaction buffer.
100 mM sodium phosphate pH 8.0.
100 mM D-phenylalanine.
100 mM L-phenylalanine.
B. Substrate and inhibitor solutions
# | [S] µM | Substrate (µL) | [Inhibitor] mM | NaP (µL) | (L)-Phe (µL) | (D)-Phe (µL) | Enzyme (µL) | Buffer (µL) | Total (µL) |
1 | 1000 | 0 | - | - | - | 100 | 1000 | ||
2 | 100 | 0 | - | - | - | 100 | 1000 | ||
3 | 25 | 0 | - | - | - | 100 | 1000 | ||
4 | 1000 | 1 | - | - | 100 | 1000 | |||
5 | 100 | 1 | - | - | 100 | 1000 | |||
6 | 25 | 1 | - | - | 100 | 1000 | |||
7 | 1000 | 0.5 | - | - | 100 | 1000 | |||
8 | 100 | 0.5 | - | - | 100 | 1000 | |||
9 | 25 | 0.5 | - | - | 100 | 1000 | |||
10 | 1000 | 10 | - | - | 100 | 1000 | |||
11 | 100 | 10 | - | - | 100 | 1000 | |||
12 | 25 | 10 | - | - | 100 | 1000 | |||
13 | 1000 | 5 | - | - | 100 | 1000 | |||
14 | 100 | 5 | - | - | 100 | 1000 | |||
15 | 25 | 5 | - | - | 100 | 1000 | |||
16 | 1000 | 10 | - | - | 100 | 1000 | |||
17 | 25 | 10 | - | - | 100 | 1000 |