[Histonet] RE: mouse brain fixation Attn: Annette Featherstone

[gayle callis]

Annette, 

 

You did not say what immunos e.g. antigens you are trying to stain?   Nor
how you are doing the actual staining method?  More information would help
please. 

 

If you are trying to stain for murine CD markers or some other cellular
antigens, there are not many that work after FFPE.  If you need to know this
information, go to SEROTEC and look at murine CD or other cellular markers,
and see if the antibody will work with FFPE paraffin applications.   BD
Bioscience also has applications that work, tested, reported, IHC or frozen
sections and found in their Technical Data Sheets for any given antibody.  

There are some other fixatives to try for CD and other cellular markers -
SEROTEC often refers to PLP (paraformaldehyde-lysine-periodate with recipe
found on web through IHCWorld or Immunoportal, Google the keywords) while BD
Bioscience refers to Becksteads Formalin free Zn TRIS buffer fixative. If
you need the recipe for that, I will be happy to send.  Otherwise BD
Bioscience would be happy to sell that to you but it is cheaper and easy to
make up in the lab. 

 

Gayle M. Callis

HTL(ASCP)HT,MT

Bozeman MT 

 

  

 

 

 

You wrote:  

 

We have been working with mouse brains that were fixed in 10% formalin for
one month. We are currently getting "no" staining with our immunos. Can this
really be the problem in light of antigen retrieval methods?

 

 

 

Annette Featherstone