[Histonet] Using lectin BS-1 for staining endothelial cells

[Koelling, Ray]

Gabriel,
It is always hard to come up with optimal solution to any staining problem
via internet since there are so many variables in procedure.  One you might
consider, if you haven't already, is the buffer used in your staining
diluent.  For biotinylated lectins, we always use HEPES buffer (not PBS).
Many lectins require Mg++ or Ca++ to bind optimally and such ions will
precipitate out in PBS.  In fact if you look at the Vector website, under
information for biotinylated Griffonia (their genus name for Bandieraea)
simplicifola I, it states that "binding appears to require divalent cations
such as calcium and magnesium".

Ray
Raymond Koelling
Research Scientist, Pathology
Amgen Corp.
Seattle, WA

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Gabriel
Anesetti
Sent: Monday, October 02, 2006 3:51 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Using lectin BS-1 for staining endothelial cells

Hello Histonetteers,
I am having problems with the staining of rat endothelial cells in 
paraffin sections using a biotin labeled lectin from Bandeiraea 
Simplicifolia BS-1 (from Sigma). The lectin does not appear to be 
staining any of the endothelial cells. I have tried using citrate buffer 
for antigen retrieval and DAB, DAB + NiCl or FICT to visualize the site 
of reaction. Sections are from ovary grafts and have very new blood 
vessels forming. Any information would be helpful.
 
Thanks
 
Gabriel Anesetti
Histology and Embryology Departament
School of Medicine
Montevideo, Uruguay   
ganesett <@t> fmed.edu.uy


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