Phenotypic Analysis of Human PBMC Subsets

Fresh PBMCs were stained with the following conjugated anti-human monoclonal antibodies (mAb): fluorescein isothiocyanate (FITC)–conjugated anti-CD21, energy-coupled dye–conjugated anti-CD19, allophyocyanin (APC)–conjugated anti-IgM, and phycoerythrin (PE)–Cy7–conjugated anti-CD27 (Beckman Coulter). PBMCs were also stained with PE-conjugated anti–FCRL-1, anti–FCRL-2, anti–FCRL-3, anti–FCRL-4, and anti–FCRL-5 mAb. We prepared these mAb as described previously (31 (link)–34 (link)). All of these FCRL-specific mAb were authorized as reference antibodies for FCRL proteins 1–5 (CD307a–e) in previous international human leukocyte differentiation antigens workshops (33 (link),35 (link)). We also used PE-conjugated anti–FCRL-4 mAb (clone 413D12) from BioLegend. PE-conjugated IgG1 (Beckman Coulter) was used as an isotype control. Flow cytometry was performed on a Navios flow cytometer (Beckman Coulter) and analyzed using CXP analysis (Beckman Coulter) and FlowJo software (Tree Star).

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Terrier B., Nagata S., Ise T., Rosenzwajg M., Pastan I., Klatzmann D., Saadoun D, & Cacoub P. (2014). CD21−/low Marginal Zone B Cells Highly Express Fc Receptor–like 5 Protein and Are Killed by Anti–Fc Receptor–like 5 Immunotoxins in Hepatitis C Virus–Associated Mixed Cryoglobulinemia Vasculitis. Arthritis & rheumatology (Hoboken, N.J.), 66(2), 433-443.

Publication 2014

PE)–Cy7–conjugated anti-CD27 Navios flow cytometer CXP analysis

Anti antibodies Anti igm Antibodies Cd307a Clone Flow cytometry Fluorescein Human Human leukocyte differentiation antigens Igg1 Isothiocyanate Isotype Monoclonal antibodies Phycoerythrin Proteins Tree Workshops

Corresponding Organization :

Other organizations : Sorbonne Université, Inserm, Centre National de la Recherche Scientifique, Sanford Research, University of South Dakota, National Cancer Institute

Top 5 similar protocols

Variable analysis

independent variables

Conjugated anti-human monoclonal antibodies (mAb)
FITC–conjugated anti-CD21
Energy-coupled dye–conjugated anti-CD19
APC–conjugated anti-IgM
PE–Cy7–conjugated anti-CD27
PE-conjugated anti–FCRL-1
PE-conjugated anti–FCRL-2
PE-conjugated anti–FCRL-3
PE-conjugated anti–FCRL-4
PE-conjugated anti–FCRL-5
PE-conjugated anti–FCRL-4 mAb (clone 413D12) from BioLegend
PE-conjugated IgG1 (Beckman Coulter) as an isotype control

dependent variables

Flow cytometry data

control variables

Staining of PBMCs

controls

Positive controls: Reference antibodies for FCRL proteins 1-5 (CD307a-e) from previous international human leukocyte differentiation antigens workshops
Negative control: PE-conjugated IgG1 (Beckman Coulter) as an isotype control

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