Quantibrite anti hla dr monocyte antibody

To quantify the expression of HLA-DR on monocyte surface, flow cytometry (FACSVerse^™ flow cytometer, BD Bioscience, Heidelberg, Germany) was conducted after incubating 50 μl ETDA-anti-coagulated whole blood with 20 μl anti-HLA-DR antibody (Quantibrite anti-HLA-DR/Monocyte antibody, BD Bioscience, Heidelberg, Germany) and after lysis of erythrocytes (FACS Lysing solution BD Bioscience, Heidelberg, Germany) as previously described and according to manufacturer`s instructions [18 (link)]. HLA-DR surface expression was measured as median fluorescence intensity after gating out monocytes based on CD14 expression (S10 Fig).
For quantification of HLA-DR molecules on cell surface (`counts per monocyte´), determined sample values were converted using daily measured 4-point calibration curves (Quantibrite PE Beads, BD Bioscience, Heidelberg, Germany).

Flow cytometry using a FACSCalibur flow cytometer (BD Bioscience, Heidelberg, Germany) was performed to determine HLA-DR expression on CD14⁺⁺-monocytes. According to manufacturer`s instructions, ETDA-anti-coagulated whole blood was incubated with anti-HLA-DR antibody (Quantibrite anti-HLA-DR/Monocyte antibody, BD Bioscience, Heidelberg, Germany) followed by erythrocyte lysis (FACS Lysing solution, BD Bioscience, Heidelberg, Germany). Daily measured 4-point calibration curves (Quantibrite PE Beads, BD Bioscience, Heidelberg, Germany) allowed the conversion of determined sample values to molecules of HLA-DR on monocyte surface.

Flow cytometry using a FACSVerseTM flow cytometer (BD Bioscience, Heidelberg, Germany) was conducted to quantify the expression of HLA-DR on CD14⁺-monocyte surface as previously described [30 (link)]. In particular, incubation of 50 μl ETDA-anti-coagulated whole blood with 20 μl anti-HLA-DR antibody (Quantibrite anti-HLA-DR/Monocyte antibody, BD Bioscience, Heidelberg, Germany) was followed by erythrocyte lysis (FACS Lysing solution, BD Bioscience, Heidelberg, Germany) according to manufacturer`s instructions. After identifying monocytes based on CD14 expression, surface expression of HLA-DR was measured as median fluorescence intensity (MFI). Conversion of determined MFI-values using a 4-point calibration curve (Quantibrite PE Beads, BD Bioscience, Heidelberg, Germany) allowed quantification of HLA-DR molecules on cell surface (`counts per monocyte´).