Re: [Histonet] RE: formalin blues / mouse CD31 IHC
Sorry....one more important step....we 3% hydrgen peroxide block (made fresh)
for 5 miunutes right after antigen retrieval.
08/28/2003 06:41 cc: email@example.com, firstname.lastname@example.org
AM Subject: Re: [Histonet] RE: formalin blues / mouse CD31 IHC(Document link: Angela
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
400 Morgan Lane
West Haven, CT 06516
"PALMER Jason (SVHM)"
Sent by: cc:
email@example.com Subject: [Histonet] RE: formalin blues / mouse CD31 IHC
08/28/2003 01:21 AM
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).
Bernard O'Brien Institute of Microsurgery
Date: Mon, 25 Aug 2003 10:30:55 -0700
From: Christian Franci
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
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
Thanks for your help!
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