Anti cd14 pe clone m5e2

The same cell staining reagents and the same reaction time were used for the CTC-FIND method and the RareCyte method. Specifically, cell staining was performed using a proprietary custom reagent kit developed by RareCyte. This included the following: 4′,6-diamidino-2-phenylindole (DAPI) to stain the cell nucleus) and antibodies specific for the epithelial marker CK, the epithelial marker EpCAM, and the cell surface antigen CD45/CD66b (used for staining leukocytes). We modified the assay by adding additional counter-stain reagent with the following phycoerythrin (PE)-conjugated antibodies: anti-CD14-PE (Clone M5E2, 1:200, BioLegend, San Diego, CA, USA), anti-CD34-PE (Clone 581, 1:200, BioLegend, San Diego, CA, USA), and anti CD11b-PE (Clone M1/70, 1:200, BioLegend, San Diego, CA, USA). For the CTC-FIND method, all staining was done with the slit-filter isolation unit of CTC-FIND. For the RareCyte method, all staining was done with the AutoStainer Link 48 (Agilent Technologies, Santa Clara, CA, USA) and according to manufacturer’s protocol.