Epo and receptor numbers-response Target epitope of your anti-epo receptor mAb is important. In order to detect the receptor on the surface of intact cells you will need an antibody which reacts with the extracellular domain of the epo receptor. Also since the receptor numbers are likely to be low a bright flourescent signal is best (eg two step staining with a PE fluorochrome). We've had some success using a purified anti-human EPO Receptor monoclonal antibody, (a murine IgG2b purified antibody), sold by R&D systems (cat #MAB307), which recognizes the extracellular domain of the natural human Epo R. To maximize detection of weak signals we used a PE conjugated anti-mouse second step antibody. The anti-Epo mAb we used, though marketed for Elisas and Westerns, worked reasonably well for flow. The cell line we studied is described as having on the order of 3,000-5,000 receptors per cell. Seperation of the anti-epo mAb stained peak from the control peak wasn't complete but was reasonable. Steven Arkin Mount Sinai School of Medicine
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