Guy.Oshiro at UCHSC.edu
Wed Jul 23 13:50:45 EST 1997
In article <33d360fd.23891111 at news.univie.ac.at>
a8803349 at unet.univie.ac.at (Martin Offterdinger) writes:
> I am currently using an IF system with an unlabelled primary antibody
> and a fluorochrome(oregon green) labelled sec. AB. I have to make this
> system more sensitive as I am only able to detect an overexpressed
> membrane receptor. My questions are: Would a system like prim.
> unlabelled, followed by sec. biotinylated AB followed by fluorochrome
> labelled Avidin be more sensitive than the system I am using now?
> Would an expected increase of a factor 10 in sensitivity be
I think that you can expect about a 4-fold increase in sensitivity by
the above approach. Another approach that you should look into is the
use of a fluorescent substrate for an enzyme linked reaction. Amersham
sells an ECF kit (enhanced Chemi Fluorescence) that basically couples
Alk Phos to the secondary and the AP acts on a fluorescence substrate
(called AttoPhos). I am planning to test that particular system out in
our lab this week - so I don't have any real data to share with you.
EMAIL: Guy.Oshiro at UCHSC.edu
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