[Histonet] lectins


Hello everyone in histoland!

I finally need to utilze the expertise of all of you fellow histonetters. 
I have limited experience in immunohistochemistry (give me a protocol and 
show me the reagents and I can follow).    I am now in a position where I 
will have to work out protocols on my own (oh no!  I will have to think!)  
Here is the first assignment.    I need to work up a protocol using 
isolectin B4 as a marker for endothelial cells in rat tissue.  Would one 
follow the application of the isolectin with an ABC reagent or, as someone 
suggested to me, use a streptavidin derivitive?  Does one need to begin 
with an antigen retreivel?  I was told that working with lectins can be 
tricky and are considered by some to be "old school" methods.   Are there 
better, newer  markers or methods for endothelial cells in rat tissue? I 
also have a reference paper that states that 100% methanol was used as a 
fixative.  Why use 100% methanol?   Is 24 hr fixation in NBF acceptable 
(or preferred)?   Any tips, comments, opinions, protocol sharing to lead 
me down the successful lectin path would be appreciated. 

Thanks and
Happy Friday to all, not much venting 
today............................well .............. those people who walk 
in from other labs and use your "stuff" and reagents 'cause they ain't got 
any can be kinda irritating...........................time for a training 
lesson...........................stay out of my kitchen and return my 

Dolores Fischer


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