RE: [Histonet] Which Collagen type for IHC in mouse tissue

From:"Jack England"

Wow.  That was some response. Thanks, all!

Per some of the questions asked, I have already done staining on my 
sections: Russell-Movat pentachrome shows my "collagen" structures to be 
yellow (which it should be), Verhoeff-van Gieson shows them to be red, and 
picro-sirius red shows them to be red as well.  When viewed with crossed 
linear polarizers (unsolicited shout-out to Edmund Industrial Optics for 
their polarizer experimenter's kit!!), these structures are refractive, 
though I don't consider my knowledge thorough enough to call this 
"birefringence" or not.  SEM images show my structures to be fibrous, and 
60-70nm thick.  All of this taken together indicates to me that I am looking 
at collagen (especially as my cells are ECM-secreting fibroblasts).

Where it has gotten tricky is in trying to figure out what flavor of 
collagen I'm looking at.  I am specifically interested in probing for 
collagens I, III, and IV, though it wouldn't be a bad idea to look at 
procollagen as well.

At the moment, though, cash is an issue, so I'm looking chiefly for collagen 
I and III antibodies that react with murine tissue.  Preferably antibodies 
that don't cross-react with other subtypes, but I'm not picky at this point.

Thanks for all the suggestions...not sure what I'm going to do for a 
collagen III antibody, but I'm going to try a rabbit polyclonal 
anti-collagen I to see if that will work.  In the meantime, I'm going to try 
to see if 1) I can sucker someone in the area out of some time with a 
cryostat, and 2) I can get ahold of a mouse tail for a positive control.

Thanks again for all your help, and best wishes!


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

_________________________________________________________________
Donít just search. Find. Check out the new MSN Search! 
http://search.msn.click-url.com/go/onm00200636ave/direct/01/


_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


<< Previous Message | Next Message >>