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:
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.
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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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