The commercially available antibodies to the following antigens were used: (i) from eBioscience, CD3 fluorescein isothiocyanate (FITC), CD8 phycoerytherin (PE) and cyanin 5.5 PE (Cy5.5PE), CD14 Cy7PE, CD16 Cy7PE, CD5 allophycocyanin (APC), CD19 Cy7PE and APC, CD20 FITC, CD45 PE, CD10 Cy7PE, CD34 Cy7PE, and tumor necrosis factor–α Alexa Fluor 700; (ii) from BioLegend, CD4 Brilliant Violet 605 (BV605) and BV785, CD8 Cy5PE, CD45RO BV570, and IFN-γ BV570; (iii) from Becton Dickinson, CD14 V500, CD3 BV605, IL-2 CF594PE, CD107a Cy5PE, granulocyte-macrophage colony-stimulating factor BV421, and macrophage inflammatory protein (MIP)–1β PE; (iv) from Beckman Coulter, CD27 Cy7PE; (v) from R&D systems, MIP-1α FITC. Alexa Fluor 647–conjugated mAb to detect the CAR19 molecule was described (32 (link)). All antibodies used in this study were titrated before use on the respective flow cytometer (see below). Routine assessment of CTL019 expansion and persistence and of CLL cells with a four-color, six-parameter Accuri C6 flow cytometer was performed as described previously (8 ).
Cd3 fluorescein isothiocyanate fitc
Manufactured by Thermo Fisher Scientific
CD3 fluorescein isothiocyanate (FITC) is a fluorescent labeling agent used in flow cytometry applications. It binds to the CD3 antigen present on the surface of T cells, allowing for the identification and enumeration of T cell populations in samples.
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Lab products found in correlation
Cd45 pe, by BioLegend (1 mentions)
Alexa fluor 700, by BioLegend (1 mentions)
Cd20 fitc, by BioLegend (1 mentions)
Cd3 bv605, by BD (1 mentions)
Mip 1β pe, by Beckman Coulter (1 mentions)
Accuri c6 flow cytometer, by BD (1 mentions)
Cd5 percp cy5.5 53 7 3, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 647 conjugated anti rat antibody, by Jackson ImmunoResearch (1 mentions)
Cytofix cytoperm plus, by BD (1 mentions)
2 protocols using cd3 fluorescein isothiocyanate fitc
1
Multiparametric Flow Cytometry Analysis of CTL019 Cells
2
Comprehensive Immune Cell Profiling
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Cells from spleen or peritoneal flushes were stained with one or more of the following anti-mouse MAbs: CD3-fluorescein isothiocyanate (FITC), CD3-phycoerythrin (PE)/Cy7 (145-2C11), GL7-eFluor 450 (GL7), F4/80-FITC (BM8), CD4-eFluor 450 (RM4-5), CD44-peridinin chlorophyll protein (PerCP)-Cy5.5 (IM7), PD-1–PE (J43), IgM-PE-Cy7 (11/41), and CD5-PerCP-Cy5.5 (53-7.3) from EBioscience; B22 0PE (RA3-6B2), CD95-PE-Cy7 (Jo2), CD19-allophycocyanin (APC)-Cy7 or -APC (ID3), CD21-FITC (7G6), and CD23-PE (B3B4) from BD Pharmingen; B220BV510 (RA3-6B2) and CD138-PE (281-2) from BioLegend; and IgG2b-FITC and IgG2c-FITC (polyclonal) from Southern Biotech. Rat anti-mouse CXCR5 (551961) (BD Pharmingen) was detected with an Alexa Fluor 647-conjugated anti-rat antibody (712-606-153) from Jackson ImmunoResearch, Inc. Cells were then assessed on a CyAn ADP flow cytometer and analyzed with FlowJo 9.3.2. (TreeStar). For intracellular staining, cells were first stained for surface markers and then fixed and permeabilized with Cytofix/Cytoperm Plus (BD Biosciences).
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