[Histonet] Collagen IHC in mouse tissue--need antibody recommendations!

[Jack England]

Aloha Histonetters, and happy holidays to all!

We've been trying to do immunostaining for collagen I on some murine-derived 
cell lines, with next to no success.  I'm almost certain that I've got 
collagen in there (PSR shows plenty of red, my poor-man's polarizers make 
the red stuff look birefringent, and SEM images show all kinds of fibrous 
ECM), but it's not staining for collagen I or III with the antibodies we've 
got (both from Sigma, produced in mouse ascites, not tested on mouse 
tissue).  I suspect that our antibodies don't react with mouse tissue, but 
it's possible that the fixation (we use 10% NBF) or the embedding (paraffin, 
which does involve heating the tissue) could be denaturing the antigenic 
sites.  I've tried enzymatic antigen retrieval, and it didn't work...next 
step is to get ahold of mouse tails and a cryostat and try for a positive 
control.

My question for anyone that can help is, assuming that the problem is that 
our antibodies do not react with mouse tissue, can any of you recommend 
antibodies to collagen I or III that do react with mouse tissue (preferably 
FFPE mouse tissue, since we don't have a cryostat)? I've checked the U of 
Iowa hybridoma bank and come up empty, and a Biocompare search gave me one 
hit--I'm curious if any of you out there can recommend something beyond that 
one.

Any takers?

--With warm regards and aloha,
Jack England
Tissue Genesis, Inc.
http://www.tissuegenesis.com

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