[Histonet] Re:human-on-human IHC

[Jamie E Erickson]

Hi Brett,
                     I haven't done exactly what your talking about but 
this may help. I'm sure your secondary anti-human will be a problem. 

When I did some crossreactivity studies in various species with a 
humanized antibody I used a protocol that was a Modification of the method 
of Tuson, Fung, Hierck and their colleagues (Tuson et al. 1990; Fung et 
al. 1992; Hierck et al. 1994) that involved mixing the primary and 
secondary together overnight at 4 degrees C in a tube.  the next day  I 
added gamma globulin (human) to bind unbound secondary then this mixture 
was applied to the section like a primary  antibody. I had good results in 
various species with little background  but did not have human tissue to 
try it out on.  It just one way to get around the secondary nonspecific 
binding. 

If you would like more information contact me I think I have the protocol.

_______________________________
Jamie Erickson
Sr. Research Associate 
Department: DSMP
Abbott Bioresearch Center
100 Research Drive
Worcester, MA 01605-4341
508-688-3134
FAX: 508-793-4895
e-mail: jamie.erickson <@t> abbott.com