RE: [Histonet] Which Collagen type for IHC in mouse tissue
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!
Tissue Genesis, Inc.
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