[Histonet] RE: formalin blues / mouse CD31 IHC

Thu Aug 28 05:53:17 CDT 2003

Sorry....one more important step....we 3% hydrgen peroxide block (made fresh)
for 5 miunutes right after antigen retrieval.

                      Angela McNabola                                                                                                  
                                               To:                                                                                     
                      08/28/2003 06:41         cc:      histonet <@t> lists.utsouthwestern.edu, histonet-admin <@t> lists.utsouthwestern.edu     
                      AM                       Subject: Re: [Histonet] RE: formalin blues / mouse CD31 IHC(Document link: 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>                                           
                      Sent by:                            cc:                                                                                     
                      histonet-admin <@t> lists.utsouth        Subject:  [Histonet] RE: formalin blues / mouse CD31 IHC                                
                      western.edu                                                                                                                 

                      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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