Legendplex th cytokine panel

Manufactured by BioLegend

The LEGENDplex Th cytokine panel is a multiplex bead-based assay for the simultaneous quantification of multiple cytokines involved in T helper (Th) cell responses. The panel allows for the measurement of cytokine levels in a single sample, providing a comprehensive analysis of the Th cell cytokine profile.

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Lab products found in correlation

Fortessa, by BD (1 mentions) Golgiplug, by BD (1 mentions) Phorbol myristate acetate (pma), by Merck Group (1 mentions) Ionomycin, by Merck Group (1 mentions) Lipopolysaccharides (lps), by Merck Group (1 mentions) Legendplex human inflammation panel, by BioLegend (1 mentions) Poly 1 c, by Merck Group (1 mentions) Anti human cd3, by Thermo Fisher Scientific (1 mentions) Anti human cd3 fitc, by BD (1 mentions) Pe anti human cd123, by BD (1 mentions) Apc cy7 anti human hla dr, by BioLegend (1 mentions) Human t cell nucleofection kit, by Lonza (1 mentions) Pan dc enrichment kit, by Miltenyi Biotec (1 mentions) Bv421 anti human cd11c, by BioLegend (1 mentions)

Close spelling variants of this product

LEGENDplex (292 citations) LEGENDplex panels (5 citations)

5 protocols using legendplex th cytokine panel

Cytokine Profiling in Serum and Supernatant

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Serum and supernatent cytokine were measured by cytokine bead array using the LEGENDPlex Th cytokine panel according to the manufacturer’s instructions (Biolegend) on a BD Fortessa flow cytometer (BD Biosciences).

Puleston D.J., Baixauli F., Sanin D.E., Edwards-Hicks J., Villa M., Kabat A.M., Kamiński M.M., Stanckzak M., Weiss H.J., Grzes K.M., Piletic K., Field C.S., Corrado M., Haessler F., Wang C., Musa Y., Schimmelpfennig L., Flachsmann L., Mittler G., Yosef N., Kuchroo V.K., Buescher J.M., Balabanov S., Pearce E.J., Green D.R, & Pearce E.L. (2021). Polyamine metabolism is a central determinant of helper T cell lineage fidelity. Cell, 184(16), 4186-4202.e20.

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Intracellular Cytokine and Activation Assays

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For detection of intracellular cytokines, cells after coculture were activated with PMA (50 ng/ml; Sigma) and ionomycin (1 μM;Merk) for 6 h in the presence of GolgiPlug (1 μl/ml BD Biosciences) for the final 4 h. Cytokines in coculture supernatants were analyzed by LEGENDplex Th cytokine panel (BioLegend).
For activation of eTAC, cDC and pDC, purified subsets were cultured at 5 × 103 cells per well in the presence of LPS (100 ng/ml; Sigma), poly(I:C) (25 μg/ml;Sigma) and R848 (25 μg/ml; Invivogen), or with PMA (50 ng/ml; Sigma) and ionomycin (1 μM;Merk). Culture supernatants were harvested after 24 h and analyzed by a custom LEGENDPlex human inflammation panel (BioLegend).

Fergusson J.R., Morgan M.D., Bruchard M., Huitema L., Heesters B.A., van Unen V., van Hamburg J.P., van der Wel N.N., Picavet D., Koning F., Tas S.W., Anderson M.S., Marioni J.C., Holländer G.A, & Spits H. (2019). Maturing Human CD127+ CCR7+ PDL1+ Dendritic Cells Express AIRE in the Absence of Tissue Restricted Antigens. Frontiers in Immunology, 9, 2902.

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Multiplex Cytokine Profiling in BAL

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Cytokines were quantified in undiluted BAL using a 13-plex cytometric bead array according to the manufacturer’s instructions (LEGENDplex™ Th-cytokine panel, BioLegend). The following cytokines were analyzed (detection limits): IL-2 (2.22 pg/ml), IL-4 (1.34 pg/ml), IL-5 (4.07 pg/ml), IL-6 (0.69 pg/ml), IL-9 (1.22 pg/ml), IL-10 (6.65 pg/ml), IL-13 (1.70 pg/ml), IL-17A (2.14 pg/ml), IL-17F (1.85 pg/ml), IL-21 (1.72 pg/ml), IL-22 (2.15 pg/ml), IFN-γ (1.39 pg/ml), TNF-α (2.09 pg/ml).

Jorde I., Hildebrand C.B., Kershaw O., Lücke E., Stegemann-Koniszewski S, & Schreiber J. (2020). Modulation of Allergic Sensitization and Allergic Inflammation by Staphylococcus aureus Enterotoxin B in an Ovalbumin Mouse Model. Frontiers in Immunology, 11, 592186.

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Multiplex Cytokine Quantification in BAL

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Cytokines were quantified in undiluted BAL supernatant in duplicates using a 12-plex cytometric bead array according to the manufacturer´s instructions (LEGENDplex™ Th cytokine panel, BioLegend). IFN-γ (0.56 pg/mL), IL-5 (1.72 pg/mL), TNF-α (2.63 pg/mL), IL-2 (1.03 pg/mL), IL-6 (2.46 pg/mL), IL-4 (0.71 pg/mL), IL-10 (10.523 pg/mL), IL-9 (1.60 pg/mL), IL-17A (0.70 pg/mL), IL-17F (1.47 pg/mL), IL-22 (3.57 pg/mL) and IL-13 (1.05 pg/mL) were analyzed (detection limits). Values that were below the detection limit were evaluated as zero.

Camp B., Jorde I., Sittel F., Pausder A., Jeron A., Bruder D., Schreiber J, & Stegemann-Koniszewski S. (2024). Comprehensive analysis of lung macrophages and dendritic cells in two murine models of allergic airway inflammation reveals model- and subset-specific accumulation and phenotypic alterations. Frontiers in Immunology, 15, 1374670.

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Silencing CEBPB in T Cells Modulates DC-Mediated Immune Responses

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DCs were enriched from PBMCs using the Pan-DC enrichment kit (Miltenyi, 130-100-777), and enriched DCs were stained with BV421 anti-human CD11c (BioLegend, 301628, 5 μl per sample), APC Cy7 anti-human HLA-DR (BioLegend, 307618, 5 μl per sample), FITC anti-human CD3 (BD Biosciences, 555232, 10 μl per sample) and PE anti-human CD123 (BD Biosciences, 340545, 5 μl per sample) antibodies. T cells were sorted from the non-DC fraction as CD3⁺ cells. DCs sorted as CD3⁻CD11c⁺HLA-DR⁺CD123⁻ were silenced and 48 h cells were stimulated with LPS (100 ng ml⁻¹) and 10 μg ml⁻¹ of OVA 323-339 (SP-51023-1, Genemed) in the presence of propyzamide (10 μM) or vehicle. After 6 h, cells were washed and co-cultured with allogenic T cells at a 1:10 ratio for 48 h. Cytokines were quantified in the supernatants using the LEGENDplex Th cytokine panel (BioLegend, 741027). CEBPB was silenced on T cells using the Human T cell nucleofection kit (Lonza, VPA-1002) and the Accell Human CEBPB-siRNA smart pool (Dharmacon, E-006423-00-0005). T cells were then activated by plate-bound anti-human CD3 (Thermo Fisher Scientific, 16-0037-85) and soluble anti-human CD28 (Thermo Fisher Scientific, 16-0289-85) in presence of propyzamide (10 μM) or vehicle.

Sanmarco L.M., Chao C.C., Wang Y.C., Kenison J.E., Li Z., Rone J.M., Rejano-Gordillo C.M., Polonio C.M., Gutierrez-Vazquez C., Piester G., Plasencia A., Li L., Giovannoni F., Lee H.G., Akl C.F., Wheeler M.A., Mascanfroni I., Jaronen M., Alsuwailm M., Hewson P., Yeste A., Andersen B.M., Franks D.G., Huang C.J., Ekwudo M., Tjon E.C., Rothhammer V., Takenaka M., de Lima K.A., Linnerbauer M., Guo L., Covacu R., Queva H., Fonseca-Castro P.H., Bladi M.A., Cox L.M., Hodgetts K.J., Hahn M.E., Mildner A., Korzenik J., Hauser R., Snapper S.B, & Quintana F.J. (2022). Identification of environmental factors that promote intestinal inflammation. Nature, 611(7937), 801-809.

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