Blood naive CD4+ T cells (75,000 cells) were cultured with anti-CD3/CD28 beads (2 µl/well) for 5 d in X-VIVO 15 serum-free medium, in the presence or absence of recombinant human IL-12p70 (2 ng/ml; R&D Bio-techne), activin A (100 ng/ml; R&D Bio-techne), and/or TGFβ1 (2 ng/ml; Peprotech). For blocking experiments with antibodies against IL-12p70 (clone 24910 from R&D Bio-techne or clone B-T21 from eBioscience), activin A (clone 69403; R&D Bio-techne), and TGFβ1,2,3 (clone 1D11; R&D Bio-techne), one dose of 20 µg/ml of each antibody or isotype control (mouse IgG1, clone 11711; R&D Bio-techne) was added at the start of the culture. For analysis of CXCL13 secretion, cells were washed and restimulated with anti-CD3/CD28 beads for 18 h in X-VIVO 15 serum-free medium.
Clone b t21
Manufactured by Thermo Fisher Scientific
The Clone B-T21 is a laboratory equipment product designed for specific scientific applications. It serves a core function within the research and analysis workflows. Further details about its intended use or performance specifications are not available at this time.
Automatically generated - may contain errors
Lab products found in correlation
Tgf β1, by Thermo Fisher Scientific (1 mentions)
Activin a, by Bio-Techne (1 mentions)
Recombinant human il 12p70, by Bio-Techne (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Ficoll paque plus, by GE Healthcare (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
2 protocols using clone b t21
1
Modulation of CD4+ T Cell Response
2
PBMC Isolation and Stimulation with PFA-fixed Bacteria
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Buffy Coats were produced from whole blood donations by using the Top & Bottom Extraction Bag System (Polymed Medical Devices). Peripheral blood mononuclear cells (PBMCs) were isolated from buffy coats by Ficoll® Paque PLUS density gradient centrifugation (GE Healthcare GmbH). PBMCs were rested overnight in RPMI 1640 medium (Gibco/Life Technologies) supplemented with 10% fetal bovine serum gold (PAA Laboratories), 2 mM L-glutamine, 50 units/ml penicillin and 50 μg/ml streptomycin (all Gibco/Life Technologies) at 37°C in a humid 7.5% CO2 atmosphere. 0.5 × 106 PBMCs were either left untreated or stimulated with PFA-fixed bacteria at different multiplicities of infection (MOI; bacteria per PBMC cell) for 6, 12, 17, and 20 h at 37°C. If indicated, PBMCs were treated with blocking antibodies against MR1 (26.5, BioLegend, 20 μg/ml) and/or against IL-12 p35 (clone B-T21, eBioscience, 5 μg/ml) or IL-18 (clone 126-2H, MBL International, 1:200 dilution) 1 h prior stimulation with PFA-fixed bacteria.
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