Re: CAM 5.2

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From:Phyllis Davie <pdavie@phenopath.com> (by way of Histonet)
To:HistoNet@pathology.swmed.edu
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Sharon,
  CAM5.2 is an antibody to cytokeratin 8.  It is sold by Becton Dickinson
(cat# 349205), and is classified "for in vitro diagnostic use", (which
puts it among the minority of commercial antibodies).
  Becton Dickinson Immunocytometry Systems
  2350 Qume Drove
  San Jose, CA 95131-1807, USA
  Ordering Information phone (800)223-8226
  Customer Support Center phone (800)448-2347
  www.bdfacs.com

We have been using it for years.  It works very well on formalin-fixed,
paraffin-embedded sections.  (I know it works on human and monkey, I
don't know about other species cross-reactivity.)  While the antibody
will work with no pretreatment, we get MUCH better sensitivity with the
following method:

  Heat in a steamer in 10mM citrate, pH 6.0, at 90C or greater for 20
minutes.
  Remove the citrate container from the steamer and allow the slides to
cool in the hot citrate for 20 minutes.
  Wash in water.
  Digest in Pronase (0.1% in PBS) for 10 minutes at room temperature.
(Calbiochem Behring cat# 53702, phone 858-450-9600  Fax: 858-453-3552)
  Wash in water.  Proceed with the primary antibody application, etc.


Hope that is helpful.

Phyllis Davie
pdavie@phenopath.com


>Can anyone give me any information of CAM 5.2 antibody?
>
>Thanks
>





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