Re: mouse mammary tumor Ab

From:"C.M. vander Loos" <c.m.vanderloos@amc.uva.nl> (by way of Histonet)

>Does anyone know where I could find an antibody that detects mouse mammary
>tumor cells (or human mammary tumor cells, but cross-reacts with mouse
>tissue)?
>
>Also, the doctor who collected the mouse tissue has had it sitting in
>alcohol for who knows how long.  Will this pose a problem with antigen
>detection?  I have only worked with formaldehyde-fixed or frozen tissues.
>
>Thanks.
>
>Jan Shivers
>Univ. of Minnesota
>Veterinary Diagnostic Lab - IHC
>1333 Gortner Ave.
>St. Paul, MN  55108
>612-625-1732; 612-624-7297
>shive003@tc.umn.edu

********

Since a number of anti-human cytokeratin antibodies tend to crossreact with
epithelia from other species, there is a fair chance that e.g.
anti-cytokeratin MNF116 (DAKO) will also crossreact with mouse tissue.
Solving the problem of mouse antibodies on mouse tissue, the Animal
Research Kit (DAKO) can be a great tool.

I don't think that alcohol-fixation will be a problem, if you take some
tissue thrinking for granted. Consider a 24-48 hours extra
fixation/emersion in Methacarn (60% methanol, 10% acetic acid-glacial, 30%
chloroform). However, be sure that upon tissue processing the samples does
not go into formalin. Alcohol-fixed tissue sections do not need any
pretreatment with heat-induced antigen retrieval or proteolytic enzymes
because the fixation type is non-crosslinking.

Chris





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