[Histonet] GS isolectin questions

[Tyrone Genade]

Hello,

I'm new to the list and the field of histology (busy startinga PhD).

I have a question regarding GS BS-I Isolectin B4 (Sigma). This was the
lectin used in the article by Kim et al (Neurobiology of Aging
25:491--500, 2004) for selective staining of CNS microglia. They state
that it was used at a dilution of 1:50 which is amazingly helpful in
light of the fact that no starting concentration is stated. I have
been able to contact the corresponding author who gave me the email of
the author who did the lectin work but no response...

I have purchased 1 mg of the GS isolectin which has now arrived in
powdered form. The recommendation by Sigma is to dissovle it in to
0.9% NaCl. Can anyone suggest a starting concentration from which I
can make the 1:50 dilutions or better, suggest a good working solution
concentration.

I will be staining retinal sections of fish, not mammals, so anyone
with advise pertinent to fish histology would be appreciated.

I understand that as negative control I will have treat control
sections with a lactose solution. Can anyone volunteer more
information regarding control protocols? Also, I may want to counter
stain with an antibody in frozen sections. Would the BSA blocking
solution interfer with the lectin binding. We had previously been
using tomato lectin with dismal results (it stuck to everything!).

Many thanks

Tyrone

-- 
Tyrone Genade
http://tgenade.freeshell.org
email: tgenade <@t> freeshell.org
tel: +27-84-632-1925 (c)
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