B7-H3 expression levels on the surface of human tumor cells were detected using purified mAb-J42 or J42-scFv-Fc. Goat anti-mouse IgG–FITC (Proteintech) was used to label the Fc of mAb-J42. The antibodies used to identify the phenotype of CAR-T cells included CD3-allophycocyanin (APC)-CY7, perforin-APC, CD4-PE, CD8-FITC, TIM3-BV711, and PD-1-BV605 (all purchased from BioLegend). Flow cytometry analysis was performed on a BD Fortessa flow cytometer and analyzed using FlowJo 10.6.0 software.
Cd3 allophycocyanin apc cy7
Manufactured by BioLegend
CD3-allophycocyanin (APC)-CY7 is a fluorochrome-conjugated antibody used for flow cytometry analysis. It binds to the CD3 antigen, which is a co-receptor expressed on the surface of T cells. The APC-CY7 fluorochrome combination provides a robust signal for identification and characterization of T cell populations.
Automatically generated - may contain errors
Lab products found in correlation
Cd4 pe, by BioLegend (1 mentions)
Pd 1 bv605, by BioLegend (1 mentions)
Tim 3 bv711, by BioLegend (1 mentions)
Cd8 fitc, by BioLegend (1 mentions)
Perforin apc, by BioLegend (1 mentions)
Fitc goat anti mouse igg, by Proteintech (1 mentions)
Fortessa flow cytometer, by BD (1 mentions)
Cd3 apc, by BioLegend (1 mentions)
Nk1.1 apc, by BioLegend (1 mentions)
Cd14 apc cy7, by BioLegend (1 mentions)
Cd11b fluorescein isothiocyanate fitc, by BioLegend (1 mentions)
Nkp46 pe cy7, by BioLegend (1 mentions)
Cd19 bv421, by BioLegend (1 mentions)
Maxisorp plate, by Thermo Fisher Scientific (1 mentions)
Brefeldin a, by Merck Group (1 mentions)
Lsrfortessa flow cytometer, by BD (1 mentions)
Golgistop, by BD (1 mentions)
Cytofix cytoperm, by BD (1 mentions)
Easysep mouse nk cell isolation kit, by STEMCELL (1 mentions)
Cd56 peridinin chlorophyll protein complex cy5, by BioLegend (1 mentions)
3 protocols using cd3 allophycocyanin apc cy7
1
Quantifying B7-H3 Expression in Tumor Cells
2
NK Cell-Mediated RVFV Gn Protein Immunity
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Three micrograms per milliliter RVFV Gn protein (custom; GenScript)-coated MaxiSorp plates (Thermo Fisher) were blocked with 5% BSA in PBST (0.01%) for 1 h at 37°C. MAbs were added to wells at 5 μg/ml and incubated for 2 h at 37°C. NK cells were isolated by negative selection from C57BL/6 mouse spleens using EasySep mouse NK cell isolation kit (StemCell Technologies). Purified NK cells were added at 2 × 105 cells/well in the presence of brefeldin A (Sigma-Aldrich), GolgiStop (BD), and anti-CD107a conjugated to phycoerythrin (PE) (BioLegend clone 1D4B) to wells already containing Gn/MAb. NK cells were incubated for 5 h at 37°C. Cells were then washed and stained with near-infrared (IR) fluorescent reactive dye (Thermo Fisher). Cells were stained for cell surface markers CD3 allophycocyanin (APC)-Cy7 (BioLegend clone 17A2), CD11b fluorescein isothiocyanate (FITC) (BioLegend clone M1/70), and NK1.1 APC (BioLegend clone PK136). The purity of NK cells was confirmed by CD3 APC (BioLegend clone 17A2), CD19 BV421 (BioLegend clone 6D5), NKp46 PE-Cy7 (Biolegend clone 29A1.4), and CD14 APC-Cy7 (BioLegend clone Sa14-2) staining. All cells were fixed in BD Cytofix/Cytoperm and then analyzed by flow cytometry on a BD LSRFortessa flow cytometer. All flow cytometric data were analyzed using FlowJo 10.7.1.
3
Immunophenotyping of NK Cell Subsets
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NK cells were labeled with fluorescently conjugated human antibodies to determine licensed and unlicensed NK cell populations: CD56-peridininchlorophyll-protein complex-Cy5.5 (BioLegend, San Diego, Calif), CD3-allophycocyanin (APC)-Cy7 (BioLegend), KIR2DL1-fluorescein isothiocyanate (R&D Systems, Minneapolis, Minn), KIR2DL2/3-APC (Miltenyi Biotec, Bergisch Gladbach, Germany), and KIR3DL1-phycoerythrin (Beckman Coulter, Fullerton, Calif). Cells were stained with antibodies at concentrations according to the manufacturers' guidelines and titrated when necessary. For intracellular staining, cells were washed with staining buffer containing PBS and 5% FBS. Cells were fixed in 2% formaldehyde for 10 minutes and then washed and permeabilized with methanol on ice for 10 minutes. They were then washed with staining buffer, followed by staining for p38 mitogen-activated protein kinase (MAPK; D13E1) antibody (Cell Signaling Technologies, Danvers, Mass). Flow cytometry was performed on an LSR Fortessa (BD Biosciences, San Jose, Calif).
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