OVA-specific Germinal Center B Cell Identification

Cell staining was performed with mAbs as listed above after blocking Fc–receptors with antibody clone 2.4G2. OVA-specific germinal center (GC) B cells were identified using 5 µg/mL OVA-488 (Invitrogen). IFNγ was detected as described (25 (link)). Cells were acquired on a Cyan ADP (Beckman Coulter) or LSR-Fortessa X20 (BD Biosciences) flow cytometer, and analyzed with Flowjo package 10 (Tree Star). Live/dead cells were distinguished using Live-Dead-eFluor780 (eBioscience).

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Nadafi R., Koning J.J., Veninga H., Stachtea X.N., Konijn T., Zwiers A., Malmström A., den Haan J.M., Mebius R.E., Maccarana M, & Reijmers R.M. (2018). Dendritic Cell Migration to Skin-Draining Lymph Nodes Is Controlled by Dermatan Sulfate and Determines Adaptive Immunity Magnitude. Frontiers in Immunology, 9, 206.

Publication 2018

OVA-488 Cyan ADP LSR-Fortessa X20

Antibody B cells Cells Clone Fc receptors Germinal center Ifnγ Mabs Tree

Corresponding Organization : Amsterdam UMC Location VUmc

Other organizations : Lund University

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Variable analysis

independent variables

Fc–receptor blocking with antibody clone 2.4G2

dependent variables

Identification of OVA-specific germinal center (GC) B cells using 5 µg/mL OVA-488
Detection of IFNγ

control variables

Live/dead cells distinguished using Live-Dead-eFluor780

controls

Positive control: None specified
Negative control: None specified

Annotations

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