Peripheral blood mononuclear cells were harvested and stained with the following antibodies according to the manufacturer's protocols: PerCP-conjugated anti-human CD4 (clone: 61D3, BioLegend), PE-CY7-conjugated anti-human CD25 (clone: M-A251, Biosciences, USA), APC-conjugated anti-human Foxp3 (clone: PCH101, eBioscience), and PE-conjugated anti-Helios (clone: 22F6, eBioscience). Corresponding isotype antibodies were used. According to the manufacturer's instructions, fixation and permeabilization were carried out after the staining of cell surface antigens CD4 and CD25, for the subsequent intracellular staining of FOXP3 and Helios. The cells were tested by flow cytometry with a fluorescence-activated cell sorter (BD Biosciences, USA). The results were analyzed with FlowJo v.X.0.7 (Treestar Inc., USA). The calculation formula [37 (link)] for the estimated absolute numbers of different cell subsets was listed in Table 1 .
Apc conjugated anti human foxp3
Manufactured by Thermo Fisher Scientific
Sourced in United States
The APC-conjugated anti-human FoxP3 is a monoclonal antibody that binds to the FoxP3 transcription factor, which is a key regulator of the development and function of regulatory T cells. The antibody is conjugated to the fluorescent dye APC (allophycocyanin), allowing for the detection and analysis of FoxP3-expressing cells using flow cytometry.
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Lab products found in correlation
Apc h7 conjugated anti human cd8, by BD (4 mentions)
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Brilliant violet bv421 conjugated anti human pd l1, by BD (2 mentions)
Bv510 conjugated anti human cd3, by BD (2 mentions)
Fluorescein isothiocyanate fitc conjugated anti human cd4, by BD (2 mentions)
Phycoerythrin pe conjugated anti human pd 1, by Beckman Coulter (2 mentions)
Pe conjugated anti human gal 9, by BioLegend (2 mentions)
Peridinin chlorophyll protein percp conjugated anti human cd56, by BD (2 mentions)
Pe conjugated anti human tim 3, by R&D Systems (2 mentions)
Fitc conjugated anti human cd4, by BD (2 mentions)
Fitc conjugated anti human cd107a, by BD (2 mentions)
Allophycocyanin apc conjugated anti human cd56, by BD (2 mentions)
Fluorescence activated cell sorter, by BD (1 mentions)
Apc conjugated anti human cd56, by BD (1 mentions)
Fitc conjugated anti human cd14, by BD (1 mentions)
Fitc conjugated anti human cd8, by BD (1 mentions)
Fitc conjugated anti human cd3, by BD (1 mentions)
Apc conjugated anti human cd8, by BD (1 mentions)
Fitc conjugated anti human cd4, by BioLegend (1 mentions)
Facscanto system, by BD (1 mentions)
7 protocols using apc conjugated anti human foxp3
1
Comprehensive Characterization of Regulatory T Cells
2
Flow Cytometric Immunophenotyping of PBMCs
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Freshly thawed PBMC were used for surface and intracellular staining regarding flow cytometric analysis. The following monoclonal antibodies were used: Brilliant Violet (BV421)-conjugated anti-human PD-L1 (BD Biosciences), BV510-conjugated anti-human CD3 (BD Biosciences), fluorescein isothiocyanate (FITC)-conjugated anti-human CD4 (BD Biosciences), FITC-conjugated anti-human CD14 (BD Biosciences), (phycoerythrin (PE)-conjugated anti-human PD-1 (Beckmann-Coulter), PE-conjugated anti-human Gal-9 (Biolegend), Peridinin Chlorophyll Protein (PerCP)-conjugated anti-human CD56 (BD Biosciences), PerCP-conjugated anti-human Perforin (BD Biosciences), allophycocyanin (APC)-conjugated anti-human TIM-3 (R&D Systems), APC-conjugated anti-human CD56 (BD Biosciences), APC-conjugated anti-human FoxP3 (eBioscience), APC/H7-conjugated anti-human CD8 (BD Biosciences).
3
Multiparametric Flow Cytometry of PBMCs
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Freshly thawed PBMC and PTL samples were used for surface and intracellular immuno-labeling and flow cytometric analysis. The following antibodies were used: fluorescein isothiocyanate (FITC)-conjugated anti-human CD4 (Clone: RPA-T4, BD Biosciences, Franklin Lakes, NJ, USA), phycoerythrin (PE)-conjugated anti-human Gal-9 (Clone: 9M1–3, Biolegend, San Diego, CA, USA), PE-conjugated anti-human TIM-3 (Clone: 344823, R&D Systems, Minneapolis, MN, USA), Peridinin-chlorophyll protein (PerCP)-conjugated anti-human CD56 (Clone: B159, BD Biosciences Franklin Lakes, NJ, USA), allophycocyanin (APC)-conjugated anti-human TIM-3 (Clone: 344823, R&D Systems, Minneapolis, MN, USA), APC-conjugated anti-human FoxP3 (Clone: 236A/E7, eBioscience, Santa Clara, CA, USA), APC-H7 conjugated anti-human CD8 (Clone: SK1, BD Biosciences, Franklin Lakes, NJ, USA) and Brilliant Violet (BV) 510-conjugated anti-human CD3 (Clone: UCHT1, BD Biosciences, Franklin Lakes, NJ, USA). Control antibodies included isotype-matched FITC-, PE-, APC-, APC-H7, or BV510-conjugated mouse antibodies.
4
Comprehensive Immune Profiling of PBMC Samples
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Thawed PBMC were used for surface and intracellular staining and analysis. The following monoclonal antibodies were used: Brilliant Violet (BV421)-conjugated anti-human PD-L1 (BD Biosciences, San Diego, CA, USA), BV510-conjugated anti-human CD3 (BD Biosciences, San Diego, CA, USA), fluorescein isothiocyanate (FITC)-conjugated anti-human CD4 (BD Biosciences, San Diego, CA, USA), FITC-conjugated anti-human CD107a (BD Biosciences, San Diego, CA, USA), phycoerythrin (PE)-conjugated anti-human PD-1 (Beckmann-Coulter, Miami, FL, USA), PE/Cy7-conjugated anti-human NKG2D (BD Biosciences, San Diego, CA, USA), allophycocyanin (APC)-conjugated anti-human CD56 (BD Biosciences, San Diego, CA, USA), APC-conjugated anti-human FoxP3 (eBioscience, San Diego, CA, USA), APC/H7-conjugated anti-human CD8 (BD Biosciences, San Diego, CA, USA).
5
Comprehensive Immunophenotyping of Activated PBMCs
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Freshly thawed PBMC were used for surface and intracellular staining and analysis. The following monoclonal antibodies were used: fluorescein isothiocyanate (FITC)-conjugated anti-human CD3 (BD Pharmingen, USA), FITC-conjugated anti-human CD4 (BD Pharmingen, USA), FITC-conjugated anti-human CD8 (BD Pharmingen, USA), FITC-conjugated anti-human CD107a (BD Pharmingen, USA), phycoerythrin (PE)-conjugated anti-human Galectin-9 (Biolegend, USA), PE-conjugated anti-human TIM-3 (R and D Systems, USA), allophycocyanin (APC)-conjugated anti-human CD56 (BD Pharmingen, USA), APC-conjugated anti-human CD8 (BD Pharmingen, USA), APC-conjugated anti-human TIM-3 (R and D Systems, USA) APC-conjugated anti-human FoxP3 (eBioscience, USA). Control antibodies included isotype-matched FITC-conjugated, PE-conjugated and APC-conjugated mouse antibodies (all from BD-Pharmingen, USA).
6
Quantification of Regulatory T Cells
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After co-culture for 3 days, cell suspensions were incubated with FITC-conjugated anti-human CD4 and PE-conjugated anti-human CD25 (Biolegend, San Diego, CA, USA) for 30 min at 4 °C and washed twice with 2 ml of phosphate-buffered saline (PBS) pH 7.4 containing 1 % bovine serum albumin (BSA). Intracellular staining for Foxp3 was then performed with APC-conjugated anti-human Foxp3 (eBioscience, San Diego, CA, USA) for 60 min and then washed with PBS/BSA. The supernatants were discarded and cells were resuspended in 0.2 ml PBS/BSA. Data were acquired with a FACSCanto system (Becton Dickinson, Franklin Lakes, NJ, USA) and analyzed using Flowjo software (Tree Star, Inc., Ashland, OR, USA). The expression levels of CD4, CD25 and Foxp3 were evaluated by calculating the percentage of cells expressing each protein.
7
Comprehensive Immune Cell Profiling Panel
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The following monoclonal antibodies were implemented: Brilliant Violet (BV) 421-conjugated anti-mouse PD-1 (BD Pharmingen), BV510-conjugated anti-mouse CD3 (BD Pharmingen), BV510-conjugated anti-mouse g/d (BD Pharmingen), fluorescein isothiocyanate (FITC)-conjugated anti-mouse CD4 (BD Pharmingen), FITC-conjugated anti-mouse CD8 (BD Pharmingen), FITCconjugated anti-mouse CD107a (BD Pharmingen), FITCconjugated anti-mouse CD49b (BD Pharmingen), FITC-conjugated anti-mouse g/d (BD Pharmingen), phycoerythrin (PE)-conjugated anti-mouse g/d (BD-Pharmingen), PE-conjugated anti-mouse TIM-3 (R and D Systems), PE-conjugated anti-mouse Gal-9 (Biolegend), PE-conjugated anti-mouse CD49b (BD Pharmingen), PE-conjugated anti-mouse CD8 (BD Pharmingen), peridinin chlorophyll a protein (PerCP)-conjugated anti-mouse CD3 (BD Pharmingen), PerCPconjugated anti-mouse CD45 (BD Pharmingen), PE-Cy7conjugated anti-mouse CD25 (BD Pharmingen), allophycocyanin (APC)-conjugated anti-human TIM-3 (R and D Systems), APCconjugated anti-human FoxP3 (eBioscience) and APC-H7conjugated anti-human CD8 (BD Pharmingen). Control antibodies included isotype-matched FITC-conjugated, PE-conjugated, PerCPconjugated and APC-conjugated rat antibodies.
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