Correct!

Amount of antigen used = 0.02 (ml = 20µl) x 4 (mg/ml BSA) = 0.08mg.

Concentration in cuvette = 0.04mg/ml (1ml precipitate + 1ml 0.1M NaOH).

If 1.9mg/ml gives an A280 of 1.0, then A280(Ag) = (0.04 x 1)/1.9 = 0.021.


Subtraction of this value from the total A280 of the precipitate gives the A280 for the antibody:

A280(Ab) = A280(ppt) - A280(Ag)

Then, given that the A280 for IgG is 1.0 at 0.7mg/ml, you can calculate the CONCENTRATION of antibody at equivalence (don't forget to take into account any dilutions made):

A)

A280(Ab) = 0.265 - 0.021 = 0.244

0.244 x 0.7 = 0.17mg IgG.

 

B)

A280(Ab) = 0.265 - 0.021 = 0.244

0.244 x 0.7 = 0.17mg/ml

0.17 x 2 = 0.34mg IgG.

C)

A280(Ab) = 0.265 + 0.021 = 0.286

0.286 x 0.7 = 0.2mg/ml

0.2 x 2 = 0.4mg IgG.

D)

A280(Ab) = 0.265 + 0.021 = 0.286

0.286 x 0.7 = 0.2mg/ml

 


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