Trfk5

Manufactured by BioLegend

TRFK5 is a mouse monoclonal antibody that recognizes the human T cell receptor complex. It is commonly used in flow cytometry applications to identify and characterize T cells.

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Lab products found in correlation

Ebio13a, by Thermo Fisher Scientific (1 mentions) Bromodeoxyuridine (brdu), by BD (1 mentions) Invivomab, by BioXCell (1 mentions) Antibody labeling kit, by Thermo Fisher Scientific (1 mentions) Anti siglecf, by BD (1 mentions) Anti amphiregulin, by R&D Systems (1 mentions) Anti il 13 ebio13a, by Thermo Fisher Scientific (1 mentions) Anti il 4 11b11, by BioLegend (1 mentions) Anti ccr3, by R&D Systems (1 mentions) Anti cd45ro uchl1, by BioLegend (1 mentions) Anti cd161 hp 3g10, by BioLegend (1 mentions) Anti cd62l mel 14, by BioLegend (1 mentions)

3 protocols using trfk5

Lung T Cell Proliferation Assay

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Lung tissue was digested with collagenase and DNAse followed by RBC lysis as above. T cell responsiveness was assessed in vitro by culturing 2 × 10⁶ total lung tissue cells with 10 μg HDM for 24h and proliferation was assessed by incorporation of 10 μM BrdU (BD Bioscience) added 1h after culture setup. Gated CD4⁺ T cells were stained with anti-BrdU (BU20A) antibody and BrdU labelling determined by flow cytometry. In other experiments with OT-II T cell transfer, lung cells were stimulated for 24h with 10 μg OVA_323–339 peptide (Innopep, San Diego). Production of IL-5 and IL-13 were measured by intracellular flow in gated CD4⁺ T cells, after adding GolgiStop & GolgiPlug for the last 5 hours, using antibodies for mouse IL-5 (TRFK5, Biolegend) and mouse IL-13 (eBio13A, ThermoFisher Scientific).

Gracias D.T., Sethi G.S., Mehta A.K., Miki H., Gupta R.K., Yagita H, & Croft M. (2020). Combination Blockade of OX40L and CD30L Inhibits Allergen-driven Memory Th2 Reactivity and Lung Inflammation. The Journal of allergy and clinical immunology, 147(6), 2316-2329.

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Activation of B-1 cells by BLyS

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B-1 cells from naïve WT mice were enriched from PerC using CD19⁺ (Miltenyi) and biotinylated CD23 mAb + streptavidin (Dynal) magnetic activated cell sorting (MACS). Purity of CD19⁺CD23⁻ B-1 cells was 96.0 ± 0.3% as assessed by IgM staining. Purified B-1 cells (CD19⁺23⁻) were cultured in the absence of stimulation with muMT splenocytes at a 1:1 ratio in cRPMI + 10% FCS supplemented with 50 μM β-mercaptoethanol (Sigma) and 50 ng/mL BLyS (R&D Systems) in the presence of either PDL2 monoclonal antibody (mAb) blockade (TY25, InVivo mAb, BioXCell) or control mAb (2A3, InVivo mAb, BioXCell) at 5 μg/mL concentration. For in vitro cultures, a STAT5 inhibitor (CAS 285986-31-4, Calbiochem) was used at 50 μg/mL and anti-IL-5 mAb (TRFK5, BioLegend) was used at 2.5 μg/mL. Cultures were harvested at day 2 or 4 for ELISA and flow cytometry analysis.

McKay J.T., Haro M.A., Daly C.A., Yammani R.D., Pang B., Swords W.E, & Haas K.M. (2017). PD-L2 regulates B-1 cell antibody production against phosphorylcholine through an IL-5–dependent mechanism. Journal of immunology (Baltimore, Md. : 1950), 199(6), 2020-2029.

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Identification and Analysis of Memory Th2 Cells

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Mouse memory Th2 cells were stained with anti-IL1rL1(ST2) (DIH9, BioLegend), and two subpopulations (ST2 low , ST2 high ) were purified by FACS. Memory Th2 cells were stimulated with immobilized anti-TCRb for 4 hours. Anti-Amphiregulin (206220, R&D Systems) was conjugated by ourselves using an Antibody Labeling Kit (Thermo Fisher Scientific) and used for intracellular staining of mouse samples. Anti-IL-5 (TRFK5, BioLegend), Anti-IL-4 (11B11, BioLegend) and Anti-IL-13 (eBio13A, eBioscience) were used for intracellular staining of mouse samples. Anti-Siglec-F (E50-2440, BD Biosciences), Anti-CCR3 (83101, R&D Systems), Anti-CD101 (Moushi101, eBioscience) and Anti-CD62L (MEL-14, BioLegend) were used for cell surface staining of mouse samples.
T cells from PBMC and dissociated cells from the ECRS polyps were stained with anti-CD4 (OKT4, BioLegend), anti-CD45RO (UCHL1, BioLegend), anti-CD161 (HP-3G10, BioLegend), anti-CRTH2 (BM16, BioLegend), anti-Lineage Cocktails (BioLegend) and anti-CD127 (A019D5, BioLegend). Anti-Amphiregulin (31221, R&D Systems) was conjugated by ourselves using an Antibody Labeling Kit (Thermo Fisher Scientific) and used for intracellular staining for human samples. Anti-IL-5 (TRFK5, BioLegend) and anti-IFNg (4S.B3, BioLegend) were used for intracellular staining of human samples.

Morimoto Y., Hirahara K., Kiuchi M., Wada T., Ichikawa T., Kanno T., Okano M., Kokubo K., Onodera A., Sakurai D., Okamoto Y, & Nakayama T. (2018). Amphiregulin-Producing Pathogenic Memory T Helper 2 Cells Instruct Eosinophils to Secrete Osteopontin and Facilitate Airway Fibrosis. Immunity, 49(1).

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