[Histonet] Flk2 / Flt3 / CD135 on mouse bone

[koellingr <@t> comcast.net]

Adam/his fellow grad student, 

  

I could be somewhat off not being up to date with current literature.  Sorry.  But used to look for Flk2/Flt3/CD135 in murine bone marrow and not the bone itself.  In immature hematopoeitic progenitor cells.  Along with looking in thymus, etc.  But for something like femurs, I removed the bone marrow, and there are some slick ways to do it without disrupting the core architecture too much.  Could take it out and obviously flow (cytometry) the cells.  But could also get out a fairly intact core and freeze it, or paraffin process/section it or glycol methacrylate section it and worry very little about the minute amount of trabecular bone in murine bone marrow. Depending on age, etc of course. 

  

So unless for the specific needs of the project call for looking actually for staining within the cortical bone, I'd take that problem out of the picture by getting the bone marrow out and then not having to worry  about decalcification and tape transfer and things like that.  Makes IHC staining a lot easier in my experience but  perhaps there are those out there who have worked out whole, intact mouse bone CD135 staining. 



Ray 



Ray Koelling 

PhenoPath Labs 

Seattle WA 



----- Original Message -----




From: "Adam ." <anonwums1 <@t> gmail.com> 
To: histonet <@t> lists.utsouthwestern.edu 
Sent: Monday, June 27, 2011 5:49:23 PM 
Subject: [Histonet] Flk2 / Flt3 / CD135 on mouse bone 

Hi all, 

One of my fellow graduate students is trying to perform immunofluorescence 
on PFA fixed, EDTA decalcified mouse bone for Flk2 (aka Flt3 or CD135). So 
far, he hasn't had any luck on either paraffin embedded or frozen sections 
using HIER. Has anyone done this successfully? I know the ideal way to 
perform IF on bone is using a tape-transfer system, but, alas, we don't 
quite have that working yet. 

Thanks, 
Adam 
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