Anti igd v450

Cell surface marker expression was analyzed by flow cytometry using a BD FACSLyric cytometer and data analysis was performed with the FlowJo software, both from Becton-Dickinson.
A surface staining protocol was applied to study the distribution of different cell subpopulations in peripheral blood. Briefly, 50 μL of peripheral whole blood were incubated with different combinations of fluorochrome conjugated monoclonal antibodies 20 min at room temperature (25°C). Red blood cells were lysed for 10 min with 2 mL of FACS Lysing solution (Becton Dickinson) and washed with phosphate-buffered saline (PBS) before flow cytometry analysis. Combinations of the following monoclonal antibodies were used: anti-CD3-PerCPCy5.5, anti-CD4-V500, anti-CD8-APCR700, anti-CD19-PECy7, anti-CD56-FITC, anti-CD45-APCH7, anti-CD45-V500, anti-HLA-DR-V450, anti-CD25-PECy7, anti-CD127-APC, anti-CD45RA-BV605, anti-CXCR5-BB515, anti-CXCR3-APC, anti-CCR6-PE, anti-CCR7-APCR700, anti-CD27-APC, anti-IgD-V450, anti-CD38-PE, anti-CD38-PerCPCy5.5, and anti-CD24-PE, all from Becton-Dickinson.

A surface staining protocol was applied to evaluate B-cell subpopulations and phenotypically classify CVID patients. Briefly, 50 µL of peripheral whole blood was incubated with a combination of fluorochrome-conjugated monoclonal antibodies (5 µL of each antibody) for 20 min at room temperature (25°C). Red blood cells were lysed for 10 min with 2 mL of FACS Lysing solution (Becton Dickinson) and washed with phosphate-buffered saline (PBS) before flow cytometry analysis. The combination of the following monoclonal antibodies was used to determine the distribution of B-cell subpopulations—anti-CD19-PECy7, anti-CD45-V500, anti-CD27-APC, anti-IgD-V450, and anti-CD21-BV605—all from Becton Dickinson.