I have been using a secondary antibody(F(ab')2 frag.) conjugated to
R-PE (size = 340 kDa) for quantitative immunostaining of a 27 kDa
protein.
I am wondering whether the size of the secondary will somehow limit
it's binding to the primary antibody-protein complex ? i.e As the
amount of protein per cell increases will the signal increase
proportionate to the amount of protein or will it exhibit some form of
saturation (non-linearity)?
Shyam Subramanian Univ. of Minnesota
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