[Histonet] Caspase-3-problem

[Goodpaster, Tracy A]

Hi Alonso,
 
We use mouse ovary/uterus for positive control for Cleaved Caspase-3.  We have used either high pH retrieval buffer or Trilogy from Cell Marque for antigen retrieval (solution preheated to over 95 deg) in a steamer for 15 minutes with a cool down in the buffer for another 15 minutes.  Then use a biotinylated secondary and ABC amplification.
 
I hope this is helpful to you!
Good luck!
 
Tracy Goodpaster BA, HTL (ASCP)
Experimental Histopathology, Shared Resources
Fred Hutchinson Cancer Research Center
 

________________________________

From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Alonso Martinez-Canabal
Sent: Fri 2/15/2008 11:24 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Caspase-3-problem




       Hi everyone,
    
        I have been trying to obtain a nice caspase-3-cleaved
immunohistochemistry  or immunofluorescence with frozen or vibratome sliced
brains (particularly hippocampus). I have been using the cell signalling
antibody, without a lot of success. I tried antigen retrieval with heating,
and combined heating with proteinase K digestion, with not good results.
Could you suggest me something?
 I also would like to know of any control tissue, I know that in normal
young mouse brain should be some positive cells in the dentate gyrus, but do
you kno any other structure that could be usable? Like skin, or liver, or
anything like that?
   Thank you very much for your valuable help
 Alonso

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Bell, Lynne
Sent: February-15-08 1:37 PM
To: Marshall Terry Dr, Consultant Histopathologist; Rene J Buesa;
WWmn916 <@t> aol.com; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Food and drink?

I wear contact lenses in the lab, but I also wear safety glasses all the
time.  I have been wearing lenses in histology for over thirty years and
have never had a problem.

And yes, Terry, when I cut onions while wearing contacts, my eyes do not
water.  I believe they do protect your eyes!!

Lynne


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