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? Many Thanks Dawn Cooper Dawn Cooper Department of Clinical Biochemistry Box 232 Addenbrooke's Hospital Hills Road Cambridge CB2 2QQ England tel: 01223 217337
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