Tom,

The general rule of thumb when staining with fluorescent protein 
conjugates is to bracket around 10 micrograms per mL.  When using a good 
fluorescent IgG conjugate, I found that 5 micrograms/mL was a bit dim, 
whereas 20 micrograms/mL often had a bit too much background.  This rule 
of thumb depends somewhat on the fluorophore (some yield a higher 
background, etc), but for TRITC conjugates, 10 micrograms/mL usually 
works great.  Although the molecular weight of your lectin is probably 
is a bit less than IgG, a 2-3 fold difference in molecular weight 
prabably won't make that much of a difference.  I used to use a TRITC 
conjugate of wheat germ agglutinin at 10 micrograms per mL and it 
stained beautifully.

Karen in Oregon
    


Date:          Fri, 03 Sep 1999 09:45:26 -0700
From:          Tom Kuwahara <tom@adpath.com>
Subject:       tomato lectins conjugated with TRITC
To:            Tim Morken <timcdc@hotmail.com>
Cc:            HistoNet@pathology.swmed.edu

Hi Tim:  Can you or anybody out there in Histonetland give me a working
range for lectins conjugated with TRITC for IHC work?  Sigma has no data
sheet or apparently no info they could give me.  Appreciate any numbers,
concentrations, etc.  anyone can give.  Thanks, Tom


>

-- 
******************************* 
Thomas J. Kuwahara
Senior Immunohistochemist
Advanced Pathology Systems 
3801 Sacramento St. suite 621 
San Francisco, CA 94118 
415 750 6800 x23067 tel 
415 750 2332 fax 
tom@adpath.com