Chunk #33 — Experimental Procedures — Immunostaining and FACS Analysis

Source

In Vitro Modeling of Blood-Brain Barrier with Human iPSC-Derived Endothelial Cells, Pericytes, Neurons, and Astrocytes via Notch Signaling.

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For FACS analysis, differentiated cells were stained with combinations of APC-conjugated PDGFRα (FAB1264A; BD Biosciences) and PE-conjugated KDR (CD309; Miltenyi Biotec), APC-conjugated CD31 (17-0319-42; BD Biosciences) and PE-conjugated PDGFRβ (558821; BD Biosciences), or APC-conjugated PSA-NCAM (130-093-273; Miltenyi Biotec) and FITC-conjugated CD44 (338804; BD Biosciences), then subjected to analysis using FACS AriaII (Becton Dickinson). Negative controls used were non-staining cells or cells stained with APC-conjugated anti-rabbit IgG (711-136-152; Jackson ImmunoResearch) and PE-conjugated anti-mouse IgG (115-116-072; Jackson ImmunoResearch).