Calculate the volumes of needed to make solutions of alkaline phosphatase and p-
ID: 956421 • Letter: C
Question
Calculate the volumes of needed to make solutions of alkaline phosphatase and p-nitrophenylphosphate with the given concentration and update the table below. Show one (1) sample calculation.
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.
A. Enzyme solutions
#
[Enzyme] ug/mL
Alkaline phosphatase (µL)
Buffer (µL)
Total volume (µL)
1
100
2000
2
125
2000
3
150
2000
B. Substrate solutions
#
[Substrate] µM
p-nitrophenylphosphate (µL)
Enzyme
((µL)
Buffer (µL)
Total volume (µL)
1
10
100
1000
2
25
100
1000
3
50
100
1000
4
100
100
1000
5
200
100
1000
6
500
100
1000
7
1000
100
1000
8
2000
100
1000
PLEASE HELP
#
[Enzyme] ug/mL
Alkaline phosphatase (µL)
Buffer (µL)
Total volume (µL)
1
100
2000
2
125
2000
3
150
2000
Explanation / Answer
1. Alkaline phosphatase stock - 500 ug/ml
Concentration(stock) x Volume(stock) = Concentration(final) x Volume(final)
Volume of stock = 100 ug/ml X 1 ml/ 500 = 0.2 ml
S.No [Enzyme] ug/mL Alkaline phosphatase stock Buffer (µL) Total volume (µL) 1 100 0.2 or 200 µL 1800 µL 2000 2 125 0.25 or 250µL 1750 µL 2000 3 150 0.3 or 300 µL 1700 µL 2000Related Questions
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