[Histonet] staining of negative controls with IHC

[FU,DONGTAO]

Hi,
  Ki67 staining in both human and mouse tissue in our lab works 
very well. We use steamer method 30min in Citra buffer.

Ann Dongtao Fu MD, Ph.D
Lab Manager
Molecular Pathology Core
Dept. of Pathology
Lab phone: 352-273-7752
Lab FAX: 352-273-7755
Lab address: D11-50
PO Box: 100275
1600 SW Archer Road
University of Flodrida
Gainesville, FL 32610


On Tue Nov 13 11:24:00 EST 2007, "Tarango, Mark" 
<mtarango <@t> nvcancer.org> wrote:

> You probably don't even need to get up to boiling for ki-67.  Try 
> 30
> minutes at 95 C in your pressure cooker.  Even on a "negative" 
> control
> you're bound to see something stain positive for ki-67, since 
> it's a
> proliferation marker... although it shouldn't be cytoplasmic.  
> I'm just
> guessing there's got to be a cell or two in the tissue that's 
> dividing.
> 
> 
> Mark Adam Tarango HT(ASCP)
> Histology & IHC Supervisor
> Nevada Cancer Institute
> One Breakthrough Way
> Las Vegas, NV  89135
> Direct Line (702) 822-5112
> Treo (702) 759-9229
> Fax (702) 939-7663
>   -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of 
> MaryAnn
> Dixon
> Sent: Tuesday, November 13, 2007 5:18 AM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] staining of negative controls with IHC
> 
> We are currently retrieving animal tissue in a pressure cooker 
> with a
> program of 125 degrees for 30 secs.  This protocol was taken from
> biocare but I'm sure this is for human tissue. Most of the tissue 
> is
> retrieved with Biocare's Reveal (pH 6.0).  We are receiving some
> background staining of our negative controls whereas if we did 
> not
> retrieve them, there is none.  We were staining for KI-67 which 
> is a
> nuclear stain. I am using a tonsil for the control. The negative 
> control
> showed some sporatic cytoplasmic staining on the Universal 
> negative
> control from biocare as well as deionized water. Since then we 
> have
> taken the temperature down to 115 degrees and increased the time 
> to 25
> minutes.  It seems to have decreased the cytoplasmic staining but 
> there
> is still some lingering.  Anyone out there have any ideas??? I'm 
> sure
> there is a better protocol for retrieving animal tissues. It has 
> to be a
> time and temperature thing!
>   MaryAnn Dixon
> Biological Scientist
> Anatomic Pathology
> University of Florida
> School of Veterinary Medicine
> 352-392-2235 ext 4517
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> 
> "EMF <nvcancer.org>" made the following annotations.
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