FW: [Histonet] Help identifying LS174T tumor cells

[Jim Manavis]

Jim

I've gone through something like this a few years ago. We attempted to
differentiate between host mouse cells and introduced human cancer cells. We
used the human mitochondrial marker successfully on FFPE tissue to
differentiate between the two. Initially we were using the Chemicon antibody
(Cat No. MAB1273) but then struck a snag with it, where with a particular
set of batch numbers it didn't work. We then turned to the Abcam antibody
(Cat No. ab3298) and have been using this ever since

Cheers

Jim

Jim Manavis
Laboratory Manager
Hanson Institute
Centre for Neurological Diseases
IMVS, Adelaide, SA, 5000
Australia
Phone: 61-08-8222-3668 / 0401120697
FAX: 61-08-8222 3392
email: jim.manavis <@t> imvs.sa.gov.au
Disclaimer: Not this little black duck! 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
koellingr <@t> comcast.net
Sent: Thursday, 15 May 2008 11:16 AM
To: jstaruk; histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Help identifying LS174T tumor cells

Jim,
Haven't worked on that cell line in lymph nodes but in other parts of mice,
xenografts and monolayers.  I'm assuming these are nude mice so the cells
aren't simply being destroyed by host (mouse) immune response.  I'd stay
away from cytokeratins.  CK20 can be downregulated in that line and there
are cytokeratins that endogenously mark in lymph nodes.  That line I believe
is presumed to be goblet cell associated.  I've looked at it with CEA and
villin, things other than CK20 or pan cytokeratin that might traditionally
mark colon.  Might try cdx2 besides CEA or villin.  I believe in ATTC that
line is known as a large producer of CEA and state of the art (back when I
did this) was to localize  LS174T's was radiolabelled CEA.  Never tried but
MUC antibody or even a PAS or mucin histology stain might do.

Raymond Koelling
PhenoPath Labs
Seattle, WA

-------------- Original message -------------- 
From: "jstaruk" <jstaruk <@t> masshistology.com> 

> OK, I've been working on this for over a week and finally decided I need 
> help. LS174T tumor cells (a human colon cancer cell line) were implanted 
> into mouse lymph nodes. I'm trying to immuno-stain these implanted cells. 
> I tried a pan cytokeratin antibody and a CK20 cytokeratin antibody and 
> neither worked. Has anyone here ever worked with this cell line? Any 
> suggestions on how to differentiate these cells from the normal mouse
cells? 
> 
> Thanks! 
> 
> Jim 
> 
> _____________________ 
> Jim Staruk 
> Mass Histology Service 
> www.masshistology.com 
> 
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