RE: [Histonet] RE: Ki-67 and Caspase 3

From:"patsy ruegg"



James I to recommend the rab mono ki67 from Labvision and for CC3 although I
have used the rab one from Cell Signaling and it works, I now get rab CC3
from BioCare it works as well in my hands and is much cheaper.  As Chris
says, I would use formalin fixation on frozens for these two markers.
Patsy 


Patsy Ruegg, HT(ASCP)QIHC
IHCtech
12635 Montview Blvd. #215
Aurora, CO 80045
720-859-4060
fax 720-859-4110
pruegg@ihctech.net
www.ihctech.net
www.ihcrg.org    
 


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of C.M. van der
Loos
Sent: Wednesday, April 16, 2008 12:52 PM
To: histonet@lists.utsouthwestern.edu
Cc: jdooley2008@yahoo.com
Subject: [Histonet] RE: Ki-67 and Caspase 3

James,Go for rabbit monoclonal clone SP6 (Thermo, Vector) for Ki67 antibody
that and rabbit anti-caspase-3 antibody from Cell Signalling. Both
antibodies works on many different species.Your cryostat tissue section from
fresh frozen blocks should be well dried (overnight, RT, under a fan) and
fixed in NBF for 5 min at RT. Acetone-fixation makes your positive nuclei
very blurry and diffuse. Wash 3-4x with TBS (or PBS) and start your
immuno.Cheers, ChrisChris van der Loos, PhD Dept. of Pathology Academic
Medical Center M2-230 Meibergdreef 9
NL-1105 AZ Amsterdam
The NetherlandsDate: Wed, 16 Apr 2008 07:15:19 -0700 (PDT)
From: James Dooley 
Subject: [Histonet] Ki-67 and Caspase 3
To: histonet@lists.utsouthwestern.edu

I would like to measure both proliferation and apoptosis. Can  anyone
recommend a good Ki-67 antibody of use on fresh frozen tissue to measure
proliferation and an antibody for activated caspase 3 for apoptosis on mouse
tissue. Please not fixation used if possible. I apologize for the confusion
with my previous e-mail.
Thank you,
James
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