Dear flowers, I have a practical question. We are doing three colour flow on mouse spleen and lymph node cells using a polyclonal rabbit anti-mouse followed by a goat anti-rabbit FITC. We want to combine two other markers like CD4 and CD8. What would be the best Abs order?: 1. Fc block (2G24); 2. polyclonal; 3. anti-rabbit FITC + CD4 and CD8 or should I stain for CD4 and CD8 separately? I apologise for the kind of question but we are in the middle of a never ending discussion here and some inputs from other flow cytometrist's experience will be greatly appreciated and hopefully bring our discussion to an end. Thank you in advance Corrado -------------------ooo--O..O--ooo-------------------------- Corrado M. Cilio, M.D., Ph.D. Immunology Research Unit Department of Endocrinology Malmo University Hospital Lund University S-205 02 Malmo, Sweden Tel: +46-40-332395; mobil: +46-70-4330338; Fax: +46-40-337042 e-mail: corrado.cilio@cmb.umu.se -------------------------------------------------------------- Judge a man by his questions rather than his answers. - Voltaire -
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