[Histonet] re:Abs against phosphoproteins
I agree with Andrea. Sure, fixation ( and don't forget pwax processing) may
well destroy/mask your protein irreversibly. Such that no amount of Ag
retrieval will work. Always is a "luck of the draw" situation. Also, if the
Ab is raised against a peptide, there is no guarantee that it will recognise
the native protein, either.
The only way to confirm that your pwax- negative staining is "real", would
be to use T/C cells that have been, for eg, irradiated, then fixed and
stained for your p-Histone. Then, you would make a pellet of another plate
of irradiated- fixed cells and process that to pwax. If the Ab is negative
on these cells, whatever pretreatment you do, then it ain't gonna work on
pwax sections, full stop. If you must use archival material, then try to
find another Ab, raised for the same protein. I'm sure others will make more
positive comments/ give alternative suggestions. Try asking here too....
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