In your case, if you do 2 rounds of staining and detect each primary  
individually with a different-colored secondary, you should have no  problem.  It is 
indeed more convenient if the primaries are from different  species, as you 
can detect both simultaneously with different-colored  secondaries.  This saves 
quite a bit of time.  You could also used  direct-conjugated primary 
antibodies, but the sensitivity may be reduced.
Albert  

Albert C.  Grobe, PhD
International Heart Institute, Tissue Engineering Lab
Saint  Patrick Hospital


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