Anti mouse cd45 pe monoclonal antibody clone 30 f11

Remaining murine kidney cell suspensions were added to FACS tubes (Corning, Corning, NY) and resuspended in PBS + . Samples were first labeled and analyzed as we recently described^{37 (link)}. Briefly, cells were co-stained with 0.5× CellMask Green (Thermo Fisher, Waltham, MA) and 2.5 μg/mL anti-mouse CD45-PE monoclonal antibody (clone 30-F11, BioLegend, San Diego, CA) at 37 °C for 20 minutes, washed twice with PBS + by centrifugation, and co-stained with 12.5 μM DRAQ5 (BioLegend) and 5 μg/mL 7-AAD (BD Biosciences, San Jose, CA) on ice for at least 15 minutes prior to analysis on an Accuri Flow Cytometer (BD Biosciences). Flow cytometry data was compensated and analyzed using FlowJo software (FlowJo, Ashland, OR). Compensation settings were established using the minced control that was digested for 60 min, CompBeads (3.0–3.4 μm diameter, BD Biosciences), and preparations with and without the above stains or an isotype matched, PE-conjugated monoclonal antibody (Rat IgG2b clone RTK4530, BioLegend) that was used as a control. Gates for each positive and negative subpopulation were defined using FlowJo to automatically calculate the compensation matrix. Finally, a sequential gating scheme was used to identify live and dead tissue cells from red blood cells, leukocytes, and non-cellular debris (see Supplementary Information, Fig. S5).