1
Bleed two has a bigger ab titer than bleed one because there is an extra ag-ab complex that isn't present in the first bleed.
2
One possible reason is that the ab titer in bleed one was too low and didn't reach the equivalence point with the antigen.
3
In bleed 1 there are a maximum of three ab-ag complexes. In bleed 2 there are four.
4
The antigen concentration decreased clockwise, since the antigens are diffusing radially from the point of application.
5
The bigger the diameter the lower the antigen concentration
6
If you look at circle "C" in bleed one when the dilution was 1/2 the antigen had to diffuse further away from the well.