Question

6. Accurate calculation of reagents is a key aspect that will determine your success or failure during the development of immunological assays. Clearly describe the steps and calculation to prepare the following reagents. (Please show work, i need to understand)

6c. 200ml of 70% EtOH from a stock of 95% EtOH

6d. 200 ul (no more than 500 ul) of each of the following dilutions of anti-IL-17 antibody 1:20 1:50

Answer 1

Question6c:

Answer:

M1V1 = M2V2

M1 = 95%

M2 = 70%

V1 =?

V2 = 200 ml

V1= volume required from stock solution

V1 = (70*200)/95

= 147.4 ml

Water required to add = 200-147.4 = 52.6 ml

Take 147.4 ml from stock solution and add 52.6 ml of water.

Question 6d:

Answer:

Dilution1: 1:20

Volume required from stock = total volume/dilution factor

= 200 μl/20

= 10 μl

Water required = 200-10 = 190 μl

Take 10 μl from stock and add 190 μl of deionized water.

Dilution2: 1: 50

Volume required from stock = 200 μl/50

= 4 μl

Water required = 200-4 = 196 μl

Take 4 μl from stock and add 196 μl of deionized water.