Intracellular staining of eosinophils
dlc32 at hermes.cam.ac.uk
Wed Feb 2 05:26:03 EST 2000
I am wanting to measure activated eosinophils with the monoclonal
antibodies EG1 and EG2 and have identified a method from previous
published papers , the FOG method, and wondered if any one would have any
experience with this technique?
the FOG method should permit the detection of intracellular antigens in
unseparated peripheral blood by flow cytometry. This uses paraformaldehyde
to fix the leukocytes and the detergent
n-octyl-beta-D-glycopyranoside(OG) to permeabilise them.
I am then staining with monoclonal antibodies specific for eosinophil
cationic protein EG1 and EG2 and a second FITC-conjugated mouse IgG.
However i am getting staining of Lymphocytes as well as eosinophils. I
wondered if anyone had any experience with this technique or problem
before, or could suggest steps in the method which could be causing this
problem if done wrong as i have little experience in this area?
Department of Clinical Biochemistry
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