[Histonet] No staining with monoclonal to Cytokeratins 5+8

[ALAN K MEEKER]

   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

   e-mail: [1]ameeker <@t> mail.jhmi.edu

References

   1. mailto:ameeker <@t> mail.jhmi.edu