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Has anyone out there done any intracellular staining for alpha
fetoprotein? I have thus far tried: 1. fixing with 1% formaldehyde in
PBS, followed by: 2. Cold MeOH permeabilization
The method seems to work pretty well for albumin, but not for AFP, and
at least one cell line should be positive, as it is an hepatocellular
carcinoma of human origin (Hep 3B).
The anti-AFP antibody I am using is NovoCastra's NCL-AFP mouse
monoclonal. I will call Novo to determine which epitope the
monoclonal recognizes while waiting for some suggestions on possible
methodologies for picking it up in cells (if this is possible, seeing
that it is a secreted protein!).
Many thanks!
If it is a secreted protein , have you tried using Brefeldin A or Monensin
in your system ?
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