With indirect immunofluorescence double labelling experiments
I have made good experiences using first ABs from different
species and goat polyklonals as 2nd ABs (FITC and Cy3 conjugated)
Now I have to set up a double labelling with two monoklonals
from mouse differing only in the isotype: IgG1 contra IgG2b.
There are lots of conjugated polyklonals that discriminate
IgG1 and IgG2b, what about the cross reactions?
The experiment should show the two antigens in one cell.
There are some monoklonal 2nd ABs (only one red) available.
I suppose they are much more specific but expensive and all
from rat. Could there occur other cross reactions e.g.
Rat at IgG2b binding to a RatIgG2b at MouseIgG1??
What system should I use monoclonal or polyclonal and are there
other tricks to get good results in this case?
Every comment is appreciated!
Anton J. Gamel
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