I used CD56 from NovoCastra diluted 1:20 with HIER at pH6 DAKO detection systems and brain control with good results.
Theresa Rohr wrote:
Is anyone out there using this antibody, CD56, from Cell Marque with the Dako Envision+ protocol. I am having some trouble arriving at a good dilution. Am presently at 1:100 with 20 min TR w/EDTA Buffer, 30 in antibody, 30 in polymer and 10 min DAB.
Some membranous staining but not really crisp and a whole lot of background staining. My control is hemorrhagic brain tissue, the only sizeable example of small cell I could find in our history. We tried carcinoid, astrocytoma and some autopsy material but the docs were not happy. What else is new???????? They want to see it on small cell and this brain mets case is the only sizeable tumor.
Does anyone have a better protocol with Envision+ and also is there another way for me to quench that background staining. I increased my H2O2 from 5 to 10 minutes and got no better results at all? Can I use Pepsin too?????
I would appreciate any advice. And yes, I did contact Cell Marque and am waiting for their suggestions too. But it's my peers that I trust the most!!!!!!!!!!
Thanks so much,
Theresa Rohr, Nyack Hospital
Theresa Rohr, BA, HT(ASCP)
Section Head, Histology
160 North Midland Avenue
Nyack, New York 10960
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