Re: TTF-1 and CKBeta34E

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From:Phyllis Davie <>
To:"Sanders, Julie, VHACIN" <>, "''" <>
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  I can't address the Ventana issue, but I will tell you what we do for TTF-1.
TTF-1     Dako cat# M3575
clone: 8G7G3/1      

We never buy pre-dilutes if it can be avoided, because I feel you have more control over your results if you do your own titers.  We run this TTF at 1:250 with HIER in citrate.  It works very nicely.  We use normal lung, or a previously positive lung cancer for our control.  Thyroid is also positive, but the level of antigen appears to be higher in thyroid than in lung, so I prefer to use lung as a control, figuring that any sensitivity issues will show up better there.

Regarding the CKBeta34E;  I believe you are referring to 34ßE12 cytokeratin antibody.  It was developed by my boss, Dr. Allen M. Gown, and is available from several vendors.  
     34ßE12 picks up cytokeratins 1, 5, 10 and 14.  It is positive on squamous epithelium, as you are seeing in your skin control.  (We use HIER in citrate for a pretreatment, but several pretreatment methods will work.)  
     In prostate, 34ßE12 picks up the outer layer of epithelium in normal, hyperplastic and atrophic prostate glands.  The inner layer of epithelium is negative for this antibody.  In prostate cancer, the outer layer of epithelium is lost.  Therefore, 34ßE12 can be used to look for prostate cancer in situations where histology alone is indeterminate.  The normal and benign glands will have a 34ßE12-positive outer layer, the carcinoma will not.  
               Wojno KJ and Epstein JL.  Am J Surg Pathol  19:251-60, 1995
               Kahane H et al.  Urology 45:981-6, 1995
     So, if you are using a prostate CANCER as your control, it WILL be negative.  You need to use a normal (or at least non-neoplastic) prostate as a control for 34ßE12.  If you are already doing this, and are still having problems, give me a call, and I'll see if we can figure anything else out.

Best of luck,
Phyllis Davie
Clinical Laboratory Supervisor
PhenoPath Laboratories, Seattle, WA
phone (206)374-9000

>Good Morning All,
>   We are having problems with the TTF-1 on the Ventana.  I have spoken to
>the vendor of the antibody, and we have tried everything under the
>sun...longer antigen retrieval time, using BioCares Nuclear Decloaker,
>different TTF-1 from other vendors (all prediluted) to no avail...any
>   Also, we do CKBeta34E on prostate, but prostate is not working as a
>control.  Skin works wonderfully, but our pathologists insist that we use a
>prostate control...again suggestions will be welcomed. 
>Thanks in advance for the responses,
>Julie A. Sanders, BA, HT(ASCP)
>Supervisor, Anatomic Pathology/Administrative Office
>VAMC, Cincinnati, Ohio

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