Detection of Ag binding in situ?

Stacy Ferguson sef at med.unc.edu
Mon Dec 18 22:52:46 EST 1995

Hi All-

I'm working with several strains of mice transgenic for particular
Ig heavy chain genes. Since these mice were generated to examine 
particular antigen responses, large percentages of B cells in each of 
these strains will stain (as determined by FACS analysis) with biotinylated
antigen followed by avidin-PE or FITC.

I would like to look at localization of these cells in situ and 
have had problems with staining of frozen tissue sections with the
same antigens used for the FACS analysis. I have no problems detecting
any other antigens usings standard immunohistochemical techniques but
realize that preservation of an Ab binding domain must be more 
difficult than preservation of individual determinants.

If anyone has experience with this type of analysis, I'd appreciate his
or her advice on fixation (if necessary) and reagents used. If the use
of fluorescence based methods are better, then I'm willing to switch. I'm
likely to try this sometime this week anyway but would appreciate some 
input anyway. 



sef at med.unc.edu
sferguso at midway.uchicago.edu


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