Ipsilateral draining submandibular and cervical lymph nodes were extracted from recipient mice (n = 6) of each group at the time of euthanization. The single-cell suspension from the lymph nodes was centrifuged and the final cell concentration was adjusted to 1 × 10 7 /mL. The 100-µl cell suspension was taken to the bottom of the flow tube. Surface staining was performed before intracellular staining. CD4-APC/C7 (Biolegend, clone # GK1.5), CD44-BV510 (Biolegend, clone # IM7), CD62-FITC (Biolegend, clone #MEL-14), CD152-APC (Biolegend, clone #UC10-4B9), GITR-PE/Cy7 (Biolegend, clone #DTA-1), Nrp-1-BV421 (Biolegend, clone #3E12), CD25-APC (Biolegend, clone #PC61), CD62L-FITC (Biolegend, clone #MEL-14), TIGIT-BV421 (Biolegend, clone #1G9), CD226-PE/Cy7 (Biolegend, clone #10E5), Helios-PerCP/Cy5.5 (Biolegend, clone #22F6), and Foxp3-PE (Thermo, clone# FJK-16s) were used in this study. Fluorescence-activated cell sorting (FACS) was performed using BD FACS LSRII Flow Cytometer. Fifty thousand lymphocytes for each sample were collected by FACS Diva 8.0 software and finally analyzed by Flowjo software (BD Bioscience, USA).
Helios percp cy5
Manufactured by BioLegend
The Helios-PerCP/Cy5.5 is a flow cytometry reagent that allows for the detection and analysis of cellular populations. It is a fluorochrome-conjugated antibody that binds to specific cell surface markers, enabling the identification and quantification of target cell subsets within a complex sample.
Automatically generated - may contain errors
Lab products found in correlation
Cd25 apc, by BioLegend (2 mentions)
Facs lsrii flow cytometer, by BD (1 mentions)
Cd152 apc, by BioLegend (1 mentions)
Nrp1 bv421, by BioLegend (1 mentions)
Gitr pecy7, by BioLegend (1 mentions)
Foxp3 pe, by Thermo Fisher Scientific (1 mentions)
Cd44 bv510, by BioLegend (1 mentions)
Cd62l fitc, by BioLegend (1 mentions)
Tigit bv421, by BioLegend (1 mentions)
Cd3 percp, by BioLegend (1 mentions)
Cd8 bv510, by BioLegend (1 mentions)
Cd4 fitc, by BioLegend (1 mentions)
Cd4 pe cy7, by BioLegend (1 mentions)
Cd27 apc, by BioLegend (1 mentions)
Cd127 pe, by BioLegend (1 mentions)
Cd25 bv421, by BioLegend (1 mentions)
Tcr aβ pe, by BioLegend (1 mentions)
Igd af488, by BioLegend (1 mentions)
Cd27 pe cy7, by BioLegend (1 mentions)
Cd45ra pe cy7, by BioLegend (1 mentions)
2 protocols using helios percp cy5
1
Characterization of Lymph Node Immune Cells
2
PBMC Flow Cytometric Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The following antibodies were used: CD3-PerCP (HIT3a), CD4-FITC (RPA-T4), CD8-BV510 ( RPA-T8), CD45RA-PE-Cy7 ( HI100), CD27-APC ( M-T271), TCR aβ-PE ( IP2b), TCR γδ-BV421 ( B1), CD19-APC ( HIB19), CD27-V450 ( M-T271), IgD-AF488 ( IA6-2), CD24-PE ( ML5), and CD38-PerCP ( HIT2), CD4-PE-Cy7 ( RPA-T4), CXCR5-BV421 ( J25ID4), CD25-APC ( MT271, ), CD127-PE ( A019D5, ), CCR6-PE ( G034E3, ), CD25-BV421 ( BC96, ), Helios-PerCP-cy5.5 ( 22F6, ), CD19-PerCP-Cy5.5 ( SJ25C1, ), CD27-PE-Cy7 ( MT271).
All were from Biolegend.
CD45RO-APC (UCHL 1), CD45RA-FITC (HI100), CXCR3-APC (1C6), and FOXP3-PE (PCH101), CD152-APC (BNI3), IgM-APC (G20-127) and IFN-γ-APC (4S.B3).
All were from BD Bioscicences. Th1 was defined as CD4 + CD45RA -CXCR5 -CXCR3 + CCR6 -, Th2 was defined as
The intracellular production of IFN-γ was investigated in PBMCs by flow cytometry. PBMCs (2 × 10 6 cells/ml) were either unstimulated or stimulated with PMA (50ng/ml) and Ionomycin (500ng/ml) for 5 hrs or BCG (MOI = 20) or 100 ng/ml BCG + IL-12 for 72 hrs, in 24-well plates. All the samples were treated with 1 μg/ml GolgiPlug (BD) for the last 2 or 6 hrs of culture.
The staining was performed according to manufacturers' guides. The samples were acquired on a FACSCanto II flow cytometer, and the data were analyzed using FlowJo.
All were from Biolegend.
CD45RO-APC (UCHL 1), CD45RA-FITC (HI100), CXCR3-APC (1C6), and FOXP3-PE (PCH101), CD152-APC (BNI3), IgM-APC (G20-127) and IFN-γ-APC (4S.B3).
All were from BD Bioscicences. Th1 was defined as CD4 + CD45RA -CXCR5 -CXCR3 + CCR6 -, Th2 was defined as
The intracellular production of IFN-γ was investigated in PBMCs by flow cytometry. PBMCs (2 × 10 6 cells/ml) were either unstimulated or stimulated with PMA (50ng/ml) and Ionomycin (500ng/ml) for 5 hrs or BCG (MOI = 20) or 100 ng/ml BCG + IL-12 for 72 hrs, in 24-well plates. All the samples were treated with 1 μg/ml GolgiPlug (BD) for the last 2 or 6 hrs of culture.
The staining was performed according to manufacturers' guides. The samples were acquired on a FACSCanto II flow cytometer, and the data were analyzed using FlowJo.
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