Hi Monica, I suggest you to check if your anti-CD13 is a mouse IgG2a. I always have problems of unspecific binding of this subclasse in my experiments. The way to avoid it is to incubate the cell suspention with some mouse normal serum (1-5%). Another possibility is that your settings are bringing some neutrophils to the lymphocyte gate. Did you check were (on the SSC x FSC graphs) your positive cells are located within the lymphocyte gate? Good Luck, Olindo
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