[Histonet] RE: formalin blues / mouse CD31 IHC

[Angela McNabola]

Hi,

We, too, in the past have had our issues with CD31 on FFPE tissue (mouse
xenographs) to be specific.  Thanks to a great technician in Tennessee that I
met at a DAKO training course (don't know if she is out there), shared a
protocol with me which works wonderfully!!!!

 We use the Santa Cruz goat  (NOT the rabbit) polyclonal PECAM-1 Cat # sc-1506.
We do everything on DAKO's immunostainer.

Basically here goes, we target retrieval in Dako TRS for approx 20/20,
avidin-biotin block (Vector, 10 min each) then we use Vector's goat elite kit.
We have found that the anitbody diluted at 1:750 works best for us (The person
that helped me recommended 1:400) and we do a 1 hour incubation at RT (or two 30
minute steps on the stainer.)

If you need anymor specifics, please feel free to contact me directly.

Hope this helps!

Angela McNabola MS, HT(ASCP)SLS
Bayer Helathcare
400 Morgan Lane
West Haven, CT 06516
203-812-5001


                                                                                                                                                  
                      "PALMER Jason (SVHM)"                                                                                                       
                      <Jason.PALMER <@t> svhm.org.au>          To:       <histonet <@t> lists.utsouthwestern.edu>                                           
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                      histonet-admin <@t> lists.utsouth        Subject:  [Histonet] RE: formalin blues / mouse CD31 IHC                                
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                      08/28/2003 01:21 AM                                                                                                         
                                                                                                                                                  
                                                                                                                                                  






 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 <cfranci <@t> rigel.com>
  To: <histonet <@t> lists.utsouthwestern.edu>
  Subject: [Histonet] Formalin blues

  I'm studying neo-vascularization.
  It seems that the only antibodies to CD31 (murine and Human) that I can find
  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 (for
  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








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