PBMCs were immunostained using various combinations of the following fluorescence-conjugated antibodies: CD4 (RPA-T4; BioLegend), CD25 (BC96; eBioscience), Foxp3 (PCH101; eBioscience), IL-17 (eBio64DEC17; eBioscience) and IFN-γ (4S.B3; eBioscience). The cells were also intracellularly stained with the following antibodies: IL-17, IFN- γ and Foxp3. Before intracellular staining, cells were stimulated in culture medium containing 25 ng/mL phorbol myristate acetate (Sigma-Aldrich), 250 ng/mL ionomycin (Sigma-Aldrich) and 1 μL/mL GolgiSTOP (BD Pharmingen) in an incubator with 5% CO2 at 37 °C for 4 h. Intracellular staining was conducted using an intracellular staining kit (eBioscience) according to the manufacturer’s protocol. Flow cytometric analysis was performed on a LSRFortessa (BD Pharmingen).
Cd25 bc96
Manufactured by Thermo Fisher Scientific
The CD25 (BC96;) is a laboratory product used for the detection and analysis of the CD25 marker in biological samples. It serves as a tool for researchers and scientists to study the expression and function of the CD25 protein, which is a key component of the interleukin-2 receptor and plays a role in immune cell activation and regulation.
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Lab products found in correlation
Phorbol myristate acetate (pma), by Merck Group (1 mentions)
Lsrfortessa, by BD (1 mentions)
Ionomycin, by Merck Group (1 mentions)
Golgistop, by BD (1 mentions)
Intracellular staining kit, by Thermo Fisher Scientific (1 mentions)
Ifn γ 4s b3, by Thermo Fisher Scientific (1 mentions)
Foxp3 pch101, by Thermo Fisher Scientific (1 mentions)
Ifn γ b27, by BD (1 mentions)
Cd4 sk3, by BD (1 mentions)
Cd38 hit2, by BD (1 mentions)
Cd86 fun 1, by BD (1 mentions)
Ctla 4 bni3, by BD (1 mentions)
Cd45ra hi100, by Thermo Fisher Scientific (1 mentions)
Cd80 l307.4, by BD (1 mentions)
Cd86 it2, by BioLegend (1 mentions)
Foxp3 236a e7, by Thermo Fisher Scientific (1 mentions)
2 protocols using cd25 bc96
1
Multiparameter Flow Cytometry of Activated PBMCs
2
Detailed Immunophenotyping of T-cell Subsets
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The following antibodies were used: CD80 (L307.4; BD Biosciences), CD86 (FUN-1; BD Biosciences), CD86 (IT2.2, BioLegend), CD25 (BC96; eBioscience), FoxP3 (236A/E7; eBioscience), CTLA-4 (BNI3; BD Biosciences), IFN-γ (B27; BD Biosciences), CD3 (UCHT1; BD Biosciences), CD4 (SK3; BD Biosciences), CD45RA (HI100; eBioscience), CD38 (HIT2; BD Biosciences) and HLA-DR (G46-6; BD Biosciences). For surface marker staining, cells were washed and re-suspended in 50 µl of FACS buffer (PBS and 2% Goat serum) containing antibodies conjugated with fluorochromes. Reactions were incubated for 30 min on ice. Cycling CTLA-4 was stained for 30 min at 37°C. For intracellular staining of FoxP3, a FoxP3 staining kit (eBioscences) was used according to the manufacturer’s instructions. Flow cytometry data were analysed by FlowJo (TreeStar, Ashland, Oregon, USA).
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