[Histonet] No staining with monoclonal to Cytokeratins 5+8
We have been trying to get epithelial cell staining in formalin-fixed
mouse prostate tissue sections with anti-CK5+8 mouse monoclonal
antibody (clone RCK 102) but are getting no signals at whatsoever.
So far we've tried antibodies from Fitzgerald and Abcam (same
clone). For antigen-retrieval we've tried either 20 minute Citrate
steaming or 40 minute steaming in target retrieval solution from DAKO.
Primary antibody was diluted in Ventana "Chem Mate" antibody dilution
buffer (up to 1:5) and incubated for 60 min. or overnight and
detection attempted with DAKO Envision kit which is working well for
us with other antibodies on these mouse prostate sections.
We are now going to try some protease treatments. Does anyone have any
experience with this monoclonal or any other suggestions. Our main
goal here is to have a good epithelial stain for the mouse
prostate that hits both basal and luminal cells.
Thanks in advance,
Alan Meeker, PhD
Assistant Professor of Pathology and Urology
The Johns Hopkins Medical Institutions
Department of Pathology
Division of Genitourinary Pathology
Bunting-Blaustein Cancer Research Building Room 124
1650 Orleans Street
Baltimore, MD 21231-1000
ph (410) 502-7880/502-7354
fax (410) 502-9817
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