Alex

	Here at the Royal Free we use Dako's CK 7 and Nova Castra's CK 19
without any problem. Our antigen retrieval is 20 minutes in pH 6.0 Citrate
buffer followed by 	15 SECONDS or less digetsion in trypsin. It is very
easy to over digect after microwaving so you may want to try between 10 and
15 secs digestion. Both Abs are 	diluted at 1/100. We use both ABC
and APAAP methods depending on tissue and or pathologist. If you want any
more info feel free to give me a call

	John Auld FIBMS MSc
	Biomedical Scientist 3
	Immunocytochemistry
	Department of Histopathology
	Royal Free Hospital
	Pond Street
	London NW3 2QG
	Tel 020 7794 0500 ext. 6516


> -----Original Message-----
> From:	HistoNet Server [SMTP:histonet@pathology.swmed.edu]
> Sent:	Wednesday, October 24, 2001 6:10 AM
> To:	HistoNet Server
> Subject:	Daily Digest
> 
> 
> 
> Date: 23 Oct 2001 11:41:49 -0500
> From: Alex Knisely 
> Subject: Can you make CK7 and CK19 work all the time?
> 
> We need help -- from you!
> 
> We need to be able to find good and reliable antibodies and staining
> protocols for cytokeratin 7 and cytokeratin 19 in liver.  From time to
> time, though controls stain beautifully and crisply for both antigens,
> using Dako anti-CK7 and BioMen anti-CK19, biopsy materials just don't
> stain.
> 
> Experimentation with varying periods of antigen retrieval has not been
> successful.
> 
> Fixation -- with core needle biopsy specimens of liver, processed
> overnight
> - -- should not, we think, be a problem.
> 
> We can not identify a different source of formalin in the biopsy
> containers
> stocked on the wards.
> 
> We are at present attempting to find out if biopsy tissues processed
> several weeks ago (before this most recent bout of "pale slide disease")
> now stain palely, or if they mark well (suggesting for the first that a
> shift in technique, for the second that a shift in processing is
> responsible).
> 
> But we would dearly like to know if someone out there can tell us with
> conviction, "Use this protocol and these antibodies -- I have NEVER had a
> problem with CK7, or with CK19, no matter what I do to the tissue!"
> 
> Thanks for any help and advice!
> 
> Alex Knisely
> 
> Alex Knisely, MD
> Consultant Histopathologist
> 
> alex.knisely@kcl.ac.uk
>  
> Institute of Liver Studies
> King's College Hospital
> Denmark Hill
> London  SE5 9RS  UK
>  
> +44 (0)20 - 7346 - 3125 telefax
> +44 (0)20 - 7346 - 4627 office
>