Figure 2

Screening of the most sensitive mAbs for Rae-1 detection. A. Flow chart of the streamlined screening process. B. Rae-1 expression in YAC-1 cells was confirmed using a commercial phycoerythrin (PE)-anti–Rae-1 antibody. Red dot: isotype control. Black solid: PE-anti–Rae-1 antibody. C. The streamlined approach and FACS analysis were used to screen 60 subclones of hybridomas producing anti–Rae-1 mAbs. YAC-1 cells were not stained, stained with isotype control antibody, or stained with culture containing a mixture of different groups’ anti–Rae-1 mAb–producing hybridoma cells. Flow cytometry was performed to determine whether the anti–Rae-1 mixture recognizes Rae-1 expression on YAC-1 cells. D. YAC-1 cells were not stained, stained with isotype control antibody, stained with PE-anti–Rae-1 antibody, or stained with each anti–Rae-1 mAb–conditioned medium of group 4. Flow cytometry was performed to assess Rae-1 expression levels.