Pe conjugated mouse igg

For flow cytometry analysis, the cells were washed twice with PBS, and 100-µl aliquots of the cell suspensions were protected from light and incubated with 10 µl of a PE-conjugated mouse anti-human CD11b antibody (BioLegend, San Diego, CA) for 60 min at room temperature. Isotype-matched PE-conjugated mouse IgG (BioLegend) antibody was used as a negative control. After the incubation, the samples were applied to a MACSQuant Analyzer (Miltenyi Biotec, Bergisch Gladbach, Germany). The mean fluorescence intensity (MFI) was calculated by subtracting the value of the mean fluorescence channel of the respective isotype control from the value obtained from the sample incubated with the specific antibody.

For FCM analysis, approximately 2–3×10⁵ cells were washed with PBS, and 30 μl aliquots of the cell suspensions were protected from light and incubated with 1 μl of a phycoerythrin (PE)-conjugated mouse anti-human CD11b antibody (BioLegend, San Diego, CA) for 30 min at room temperature. An isotype-matched PE-conjugated mouse IgG (BioLegend) antibody was used as a negative control. After incubation, the samples were analyzed on an LSR Fortessa X-20 Flow cytometer (BD Biosciences, San Jose, CA).