immunofluorescence: direct vs. indirect
peter.cherepanov at uz.kuleuven.ac.be
Tue Jun 23 07:15:06 EST 1998
What would be the best choice for doing immunofluorescence:
to obtain labeled monoclonal antibodies or to use
labeled secondary antibody ?
I heard that using secondary antibodies increases sensitivity -
is that true ?
(As I understand the primary antibody is usually an IgG, and
the bulk of the secondary is IgGs too, which are monomeric.
Therefore, I would expect only a drop in sensitivity and
specificity. Using biotinilated antibodies may have an advantage,
since avidine is tetrameric and can bring more FITC residues to
the spot (?))
I need not only sensitive, but also very specific detection.
(surface staining of monocytes, I plan to use CD14 antibody).
Would like to hear any hints.
More information about the Methods