Re: [Histonet] RE: formalin blues / mouse CD31 IHC

From:Gayle Callis

We use frozen sections avoiding FFPE entirely. Check Histonet Archives on
at Histosearch website.  There have been messages about many times in the
past. Check out this answer from Barb Wright. 


Re: Immunos on Mouse tissue
From: Barbara Wright  

If you are using the DAKO CD31 antibody - it will not react with mouse
endothelium.  Pharmigen CD31 (MEC13.3) works well on FFPE with Vectastain
ABC followed by NEN TSA-kit.  And our Collagen IV is from a polyclonal from
Chemicon.  Both work well on FFPE mouse tissues.
Barb Wright wrote:

> I am in need of some help? I am trying to do immunos on mouse tissue. I
> currently use the Dako LSAB+ kit and I know that some of my antibodies are
> monoclonal. I have tried the Dako ARK kit and have not had any luck. I don't
> know what I may have done wrong. I also have tried the Dako Envision
> polyclonal kit. I have changed my antibodies from mono to poly or goat. I
> have tried different retrieval methods. What can I do now? Any help would be
> appreciated. The two antibodies I am working with mostly is the Cd31 (goat
> polyclonal and Rat anti mouse) and Collagen 4 (monoclonal and polyclonal).
> Thanks
> Donella Stillings
>	Hi all.
>	I'm also trying to get an anti mouse CD31 to work on FFPE sections (CBL
1337 from Cymbus / Chemicon). Has anybody had success of any sort with this
antibody and type of tissue, and if so, what's your secret? (By the the
way, Dako anti human CD31 (M 0823) works just fine on human FFPE tissues
for me).
>	Much obliged!
>	Jason Palmer
>	Bernard O'Brien Institute of Microsurgery
>	Melbourne, Australia
>		Date: Mon, 25 Aug 2003 10:30:55 -0700
>		From: Christian Franci 
>		To: 
>		Subject: [Histonet] Formalin blues
>		I'm studying neo-vascularization.
>		It seems that the only antibodies to CD31 (murine and Human) that I can
>		commercially (BD Bio and Chemicon) do not work at all for formalin fixed,
>		paraffin imbedded tissue sections.
>		Does anyone out there know of any anti CD31 Ab's that DO work in formalin
>		fixed tissues?
>		Also, does anyone know of any Ab that I could use as a positive control
>		both mouse and human tissues) that works well in formalin-fixed samples?
>		Lastly, does anyone have a better fixation method to formalin (compatible
>		with paraffin imbedding) which may allow me to use the above mentioned
>		commercial Ab's?
>		Thanks for your help!
>		Cheers,
>		Chris

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology 
Montana State University - Bozeman
S. 19th and Lincoln St
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

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