Flow cytometric measurements were done in one study centre (Muenster). For this purpose, PB and CSF were analysed within one hour after sampling. The CSF was centrifuged and treated with VersaLyseTM (Beckman Coulter, Krefeld, Germany) in parallel to PB, according to the manufacturer’s instructions. Cells were incubated with fluorochrome-conjugated monoclonal antibodies [CD14-FITC (clone RM052), CD138-PE (clone B-A38), HLA-DR-ECD (clone Immu-357), CD3-PC5.5 (clone UCHT1), CD56-PC7 (clone N901), CD4-APC (clone 13B8.2), CD19-APC A700 (clone J3-119), CD16-APC A750 (clone 3G8), CD8-PacificBlue (clone B9.11), CD45-KromeOrange (clone J.33); all Beckman Coulter, dilution 1:200] for 30 min, washed and analysed by flow cytometry on a NaviosTM (Beckman Coulter) flow cytometer.32 (link) Data were analysed with KaluzaTM 2.1 software (Beckman Coulter; for gating strategy, see Supplementary Fig. 1 ). Routine CSF parameters were investigated in addition to flow cytometry in both study centres (Magdeburg and Muenster). Cells were counted using a Fuchs-Rosenthal chamber. The CSF/serum IgG, IgA, IgM, and albumin ratio and the blood/CSF-barrier integrity were determined by nephelometry (BN ProSpecTM, Siemens Healthcare). IgG oligoclonal band (OCB) patterns were analysed by isoelectric focussing in gel-electrophoresis and subsequent silver staining (Processor PlusTM, GE Healthcare).
Cd56 pc7 clone n901
Manufactured by Beckman Coulter
The CD56-PC7 (clone N901) is a flow cytometry reagent that recognizes the CD56 antigen. CD56 is a neural cell adhesion molecule (NCAM) expressed on natural killer (NK) cells, certain T cell subsets, and some tumor cells. This reagent can be used to identify and enumerate CD56-positive cell populations in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Navios, by Beckman Coulter (1 mentions)
Hla dr ecd clone immu 357, by Beckman Coulter (1 mentions)
Versalyse, by Beckman Coulter (1 mentions)
Live dead fixable near ir, by Thermo Fisher Scientific (1 mentions)
Violet tracer, by Thermo Fisher Scientific (1 mentions)
Cd38 pe clone hb 7, by BD (1 mentions)
Lsrfortessa, by BD (1 mentions)
2 protocols using cd56 pc7 clone n901
1
Flow Cytometry Analysis of CSF and Peripheral Blood
2
Multicolor Flow Cytometry Assay for CAR T-Cell Characterization
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Flow cytometry assays were performed on BD LSRFortessa. Viable cells were determined with live/dead cell marker (LIVE/DEAD® Fixable Near-IR; Life Technologies L10119). Transduction efficiency and associated CAR expression was measured with an monoclonal antibody towards NGFR-APC (CD271) (clone ME20.4 Biolegend). Monoclonal antibodies used for cytotoxicity assays: CD3-Fitc (clone SK7), CD14-PerCP (clone MoP9), CD19-PerCP (clone SJ25C1) and CD38-PE (clone HB7) (BD Bioscience). CD56-PC7 (clone N901) and CD138-APC (clone BA38) (Beckman Coulter). To distinguish Mock/CAR T cells from target cells, target cell were stained with 0.5 μM Violet tracer (Thermo Fisher C34571) for 25 minutes and washed before cytotoxicity assay co-cultures. Flow cytometry data analysis was performed with FACS Diva 6.1 software.
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