[Histonet] Endothelial cell staining-vWF-A word of caution

[Rocan]

Jerry,
No doubt that many histonet pros have great protocols for EC 
immunostaining.  However, there is much controversy about vWF as a good 
endothelial marker.  Please refer to this link:
ww-klinik.uni-mainz.de/Pathologie/E/ZuStaffProfiles/ 
Kirkpatrick/PDF-Dateien/1_multipart_xF8FF_14_16-Kirk.pd.
There are plenty of vascular biologist who would agree that vWF is not 
the best marker for microvessels not only in human tissue but in other 
species as well including mice (Dev Biol. 1991 Nov;148(1):51-62).
Although vWF is an excellent marker for vein endothelium, it is less so 
for arteries.   Venules and arterioles stain ok but capillary 
endothelium  has at best a weak reaction and mostly does not have 
detectable vWF.   This has much more to do with the biology of vWF than 
the quality of the antibodies. A colleague of mine had a manuscript 
rejected because he used vWF as EC marker in retina.  The reviewers 
demanded the use of CD31. Furthermore, tumors can be negative to vWF in 
much of their vasculature.  vWF can be used in addition to CD31 but 
never instead.  CD34 also can be used to complement vWF since it is 
strongest in capillaries.  However, no one would argue with a good CD31 
stain which stains all vessels regardless of their caliber.
Therefore, I think that for mouse tissue, the Pharmigen  clone MEC13.3 
(Eur J Cell Biol. 1994 Apr;63(2):247-54) is your best option.
Professor Donald McDonald from UCSF  has made a career on vascular 
imaging.  I have learned so much from reading his papers and paying 
good attention to his protocols. You may want to go to start with this 
fantastic review and then go to specific protocols in his many 
publications.  I assure you he is the best.
  McDonald DM, Choyke PL.  Imaging of angiogenesis: from microscope to 
clinic. Nat Med. 2003 Jun;9(6):713-25
Good luck

-----
Dr.Rocio Sierra-Honigmann
Director
Engineered Wound Repair Laboratory
Cedars Sinai Research Institute
Davis 1091
310-423-1882
Honigmannr <@t> cshs.org	
On Jul 23, 2004, at 8:52 AM, Jeremiah Lyons wrote:

> Hi!
>
> I'm a PhD student who is looking for advice on endothelial cell 
> staining in mouse tumour tissue for the purpose of microvessel density 
> staining.Most of the available antibodies don't work in mouse tissue. 
> At the moment I'm trying Carnoy's fixed tissue (fixed for four hours) 
> with rabbit anti-human Von Willebrand factor. Any suggestions would be 
> greatly appreciated.
>
> Jerry
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