Rea control human igg1 apc conjugated isotype antibody

Cells were trypsinized, centrifuged, and resuspended as above. For mitochondrial superoxide staining, cells were incubated with 2.5 μM MitoSOX Red (Thermo Fisher, M36008) in 100 μL PBS for 1 h room temperature, rinsed with 1x PBS, centrifuged for 3 min at 300g, and resuspended in 5% FBS in PBS without fixation. For all other flow assays, cells were fixed with 4% paraformaldehyde for 5 min. For staining of patient line iPSC-CMs with cardiac troponin T (cTnT), cells were incubated with cardiac troponin T APC-conjugated antibody (Miltenyi Biotec, 130-120-403) or REA control human IgG1 APC-conjugated isotype antibody (Miltenyi Biotec, 130-120-709) for 1 h at room temperature in the dark in 0.75% saponin (Sigma, 558255-100G) and 5% FBS in 1x PBS. Cells were rinsed with 1x PBS, centrifuged for 3 min at 300g, and resuspended in 5% FBS in 1x PBS for analysis using a FACSCanto cytometer. Populations were serially gated for FSC-A/SSC-A, FSC-H/FSC-W, and GFP+/AmCyan- to identify EGFP-expressing iPSC-CMs.