[Histonet] commercial antibody diluents w/ surfactants

[Gayle Callis]

Question:  are you planning to do immunofluorescent staining even though you 
are now doing only enzyme immunohistochemistry with these reagents?

The size of the fluorophore molecules is not a factor but there are some 
other considerations when doing IF.

Gayle M. Callis
HTL/HT/MT(ASCP0
Bozeman MT 59715



----- Original Message ----- 
From: "Brianna Mbow" <bmbow <@t> seagen.com>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Friday, April 18, 2008 11:47 AM
Subject: [Histonet] commercial antibody diluents w/ surfactants


Hello,

In my previous position, I prepared all my own antibody solutions for
fluorescent staining myself with varying levels of blocking serum and 
triton-x for
permeabilization.

I'm now working in a lab that uses commercial antibody diluents and an 
autostainer with its
own washing solution.  Both the diluent and wash solution contain 
surfactant, but the concentration is
not listed.  I'm just wondering if anyone knows if the amount is sufficient 
for permeabilization in
fluorescent staining.  So far in the lab we have only really been doing IHC 
and they have never added triton-x to
their solutions.  Also, out of general curiosity, is there a reason for 
needing more permeabilization in IF
compared to IHC staining?  Is it because of the size of fluorophore 
molecules?  Any ideas would be much
appreciated.



Thank you,

Brianna Mbow

Antibody Technologies

Seattle Genetics

Seattle, WA



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