Cells from the thymus, spleen or lymph nodes were resuspended in staining buffer at a density of 2 × 106 cells/ml. The cells were incubated with anti-FcγRII/III monoclonal antibody (mAb) (2.4G2), followed by antibodies against CD4 (RM4-5, BD Pharmingen), CD8 (53–6.7, BD Pharmingen), B220 (RA3-6B2, BioLegend), Thy1.1 (53–2.1, BioLegend), Thy1.2 (OX-7, BioLegend), CD44 (IM7, BioLegend), CD62L (MEL-14, BioLegend), Dll1 (HMD1-5, eBiosciences), Dll4 (HMD4-1, eBiosciences), Jagged1 (HMJ1-29, eBiosciences) or Jagged2 (HMJ2-1, eBiosciences). After gating out cells that were positive for 7-AAD, excluding the doublets, the fluorescence intensity of 105 cells was measured with a FACS Canto II flow cytometer (BD Biosciences, CA) and analyzed with FACSDiva (BD Biosciences) or FlowJo (Tree Star) software programs. For intracellular staining, cells were fixed with an intracellular staining kit (72–5775, eBioscience), permeabilized and stained with Foxp3 (3G3, Tonbo Biosciences) antibody.
Foxp3 3g3
Manufactured by Cytek Biosciences
Foxp3 (3G3) is a flow cytometry reagent produced by Cytek Biosciences. It is used for the detection and analysis of Foxp3-expressing cells, which are typically regulatory T cells. The reagent is designed to provide a reliable and consistent measurement of Foxp3 expression in various cell types.
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Lab products found in correlation
Facscanto 2 flow cytometer, by BD (1 mentions)
Facsdiva, by BD (1 mentions)
Intracellular staining kit, by Thermo Fisher Scientific (1 mentions)
Cd62l mel 14, by BioLegend (1 mentions)
Cd44 im7, by BioLegend (1 mentions)
Rm4 5, by BD (1 mentions)
Cd8 53 6, by BD (1 mentions)
B220 ra3 6b2, by BioLegend (1 mentions)
Facscanto 2 system, by BD (1 mentions)
Annexin 5 fitc apoptosis detection kit, by Immunostep (1 mentions)
Sytox blue, by Thermo Fisher Scientific (1 mentions)
Tcrβ h57 597, by BD (1 mentions)
Cd62l mel 14, by BD (1 mentions)
Cd3 145 2c11, by Cytek Biosciences (1 mentions)
Nur77 12.14, by Thermo Fisher Scientific (1 mentions)
2 protocols using foxp3 3g3
1
Comprehensive Immune Cell Profiling
2
Multiparametric Flow Cytometry Staining
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Cell staining was done with fluorescently labeled mAbs to CD5 (53–7.3), CD4 (RM4-5), CD8α (53–6.7), CD24 (M1/69), TCR-Vα2 (B20.1TCR-Vβ6 (RR4-7), TCRγ (GL-3), and TCRβ (H57-597) were purchased from BD; CD62L (MEL-14), CD44 (IM7), CD69 (H1.2F3), CD3 (145.2C11), and FoxP3 (3G3) were obtained from TONBO Bioscience; CD25 (PC61.5), CD19 (6D5), CD161 (PK136), and CD6 (OX129) were purchased from BioLegend; and Nur77 (12.14) was obtained from eBioscience; all were used according to the manufacturer’s instructions. FACS analyses were done on a LSRII 564 or FACS Canto II system (BD) using FlowJo software (TreeStar). Cell viability was evaluated either with SYTOX Blue (Life Technologies) or 633- or 405-nm Viability dyes (Life Technologies). Apoptotic cells were detected by using Annexin V-FITC apoptosis detection kit (Immunostep).
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