Rhil 13

Manufactured by R&D Systems

Sourced in United States

RhIL-13 is a recombinant human interleukin-13 (IL-13) protein. IL-13 is a cytokine that plays a role in immune and inflammatory responses.

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Lab products found in correlation

Rhil 4, by R&D Systems (4 mentions) Rhifn γ, by R&D Systems (4 mentions) Phorbol myristate acetate (pma), by Merck Group (2 mentions) Isotype igg, by R&D Systems (1 mentions) Rhil 6, by R&D Systems (1 mentions) Mouse anti β actin, by Beyotime (1 mentions) Phorbol 12 myristate 13 acetate, by Merck Group (1 mentions) Matrigel, by BD (1 mentions) M csf, by R&D Systems (1 mentions) Anti il 6, by R&D Systems (1 mentions) Pimozide, by Merck Group (1 mentions) Piceatannol, by Merck Group (1 mentions) Nvp bez235, by Selleck Chemicals (1 mentions) Rad001, by Chemietek (1 mentions) Annexin 5 fitc kit, by Thermo Fisher Scientific (1 mentions) Rdea119, by Selleck Chemicals (1 mentions) Rhil 5, by BD (1 mentions) Recombinant canine il 4, by R&D Systems (1 mentions) 3h thymidine, by Cytiva (1 mentions) Rhil 4, by Thermo Fisher Scientific (1 mentions)

7 protocols using rhil 13

Caco-2 Cell Cytokine Stimulation

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Caco-2 cells were grown in DMEM supplemented with 10% FBS, 100 U/ml penicillin, 100 mg/ml streptomycin and 110 mg/L pyruvate. Confluent Caco-2 cells grown on 24-well plates were fasted for 12h and then stimulated with recombinant human (rh)-IFN-γ (10ng/ml), rh-IL-4 (100 ng/ml) or rh-IL-13 (10ng/ml) (R&D Systems, Minneapolis, MN, USA) for 90 min, for further immunoblotting assays. Chemokines in cell-culture supernatants were assayed after stimulation with FliC (1µg/ml), rh-IFN-γ (10ng/ml), rh-IL-4 (100 ng/ml), rh-IL-13 (10ng/ml), rh-IL-5 (10ng/ml) (R&D Systems, Minneapolis, MN, USA), Dupilumab (human anti-interleukin-4 receptor alpha monoclonal antibody) (50µg/ml) (Sanofi Genzyme, Regeneron, NY, USA) or Mepolizumab (human anti-interleukin-5 monoclonal antibody) (50 µg/ml) (GSK, San Polo di Torrile, Italy) for 48h at 37°C. We also used a specific JAK1 and JAK2 inhibitor, Ruxolitinib (Novartis, Stein, Switzerland), at different doses for 2h. The inhibitors and monoclonal antibodies were diluted in medium and incubated with cells alone or in combination with the different stimuli.

Vaccaro J., Canziani K.E., Guzmán L., Bernedo V., García M., Altamirano E.M., Feregotti E., Curciarello R., Muglia C.I, & Docena G.H. (2022). Type-2 Cytokines Promote the Secretion of the Eosinophil–Attractant CCL26 by Intestinal Epithelial Cells in Food-Sensitized Patients. Frontiers in Immunology, 13, 909896.

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Macrophage Polarization and Activation Assay

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Isotype-IgG, rhIL-4, rhIL-13, M-CSF, rhIL-6, anti-IL-6 and human IL-6 Quantikine ELISA were purchased from R&D Systems. Phorbol 12-myristate 13-acetate (PMA) was purchased from Sigma. Bisindolylmaleimide I (protein kinase C (PKC) inhibitor) was purchased from Calbiochem. Matrigel was from BD Biosciences. The primary human antibodies in this study were used: mouse anti-CD163, rabbit anti-CD31 (ZSGB-BIO); rabbit anti-MMP14, rabbit anti-LAMC2 (Abcam); rabbit anti-MMP9, rabbit anti-Phosphorylated PKC (pan) (Cell Signaling Technology); mouse anti-β-actin (Beyotime).

Zhang L., Xu Y., Sun J., Chen W., Zhao L., Ma C., Wang Q., Sun J., Huang B., Zhang Y., Li X, & Qu X. (2016). M2-like tumor-associated macrophages drive vasculogenic mimicry through amplification of IL-6 expression in glioma cells. Oncotarget, 8(1), 819-832.

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Cytokine-Mediated Cell Proliferation Assay

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Masitinib and midostaurin (PKC412) were purchased from LC Laboratories (Woburn, MA, USA), piceatannol and pimozide from Sigma-Aldrich (St Louis, MO, USA), RDEA119, PD0325901 and NVP-BEZ235 from Selleck (Houston, TX, USA) and RAD001 from ChemieTek (Indianapolis, IN, USA). The antibody-drug conjugate brentuximab vedotin (SGN-35) was kindly provided by Dr P. Veiby and Dr J. V. Garafalo (Millennium Takeda Oncology Company, Cambridge, MA, USA). Stock solutions of drugs were prepared by dissolving in dimethyl sulfoxide (Merck, Darmstadt, Germany). Recombinant human (rh) interleukin (IL)-2 was obtained from ImmunoTools (Friesoythe, Germany), rhIL-4 from Peprotech (Rocky Hill, NJ, USA), rhIL-5 from BD Biosciences (San Jose, CA, USA), rhIL-6 from Novartis (Basel, Switzerland), rhIL-13, rhCD30 ligand, recombinant canine (rc) IL-4, and rc stem cell factor (SCF) from R&D Systems (Minneapolis, MN, USA) and rhSCF from Strathmann Biotech (Hannover, Germany). RPMI 1640 medium and fetal calf serum (FCS) were purchased from PAA Laboratories (Pasching, Austria), ³H-thymidine from Amersham (Buckinghamshire, UK) and the Annexin V-FITC Kit from eBiosciences (San Diego, CA, USA).

Bauer K., Hadzijusufovic E., Cerny-Reiterer S., Hoermann G., Reifinger M., Pirker A., Valent P, & Willmann M. (2016). IL-4 downregulates expression of the target receptor CD30 in neoplastic canine mast cells. Veterinary and comparative oncology, 15(4), 1240-1256.

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Isolation and Differentiation of Monocyte-Derived Cells

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CD14⁺ monocytes were isolated from fresh PBMCs with CD14 MicroBeads (Miltenyi Biotec, 130-050-201) according to the manufacturer’s instructions. For the induction of monocyte-derived macrophages (MDMs), CD14⁺ monocytes were cultured in RPMI 1640 supplemented with 10% FCS, 1% penicillin/streptomycin and 50 ng/ml recombinant human (rh)M-CSF (R&D Systems, 216-MC) for 7 days, and were then allowed to differentiate for seven days in the presence of 50 ng/ml rhM-CSF and 50 ng/ml rhIL-4 (R&D Systems, 204-IL). Monocyte-derived dendritic cells (MDDCs) were induced by culturing CD14⁺ monocytes for 7 days in RPMI 1640 supplemented with 10% FCS, 1% penicillin/streptomycin, 50 ng/ml rhGM-CSF (R&D Systems, 215-GM) and 20 ng/ml rhIL-13 (R&D Systems, 213-ILB). For osteoclast differentiation, CD14⁺ cells were cultured in α-MEM (Thermo Fisher Scientific, 12571063) containing 10% FCS, 1% penicillin/streptomycin, 30 ng/ml rhM-CSF and 10 ng/ml rhRANKL (Peprotech, 310-01) for 7 days. The medium was replaced every three days.

Bohlen J., Zhou Q., Philippot Q., Ogishi M., Rinchai D., Nieminen T., Seyedpour S., Parvaneh N., Rezaei N., Yazdanpanah N., Momenilandi M., Conil C., Neehus A.L., Schmidt C., Franco C.A., Le Voyer T., Khan T., Yang R., Puchan J., Erazo L., Roiuk M., Vatovec T., Janda Z., Bagaric I., Materna M., Gervais A., Li H., Rosain J., Peel J., Seeleuthner Y., Han J.E., L’Honneur A.S., Moncada-Vélez M., Martin-Fernandez M., Horesh M.E., Kochetkov T., Schmidt M., AlShehri M.A., Salo E., Harri S., ElGhazali G., Yatim A., Soudée C., Sallusto F., Ensser A., Marr N., Zhang P., Bogunovic D., Cobat A., Shahrooei M., Béziat V., Abel L., Wang X., Boisson-Dupuis S., Teleman A.A., Bustamante J., Zhang Q, & Casanova J.L. (2023). Human MCTS1-dependent translation of JAK2 is essential for IFN-γ immunity to mycobacteria. Cell, 186(23), 5114-5134.e27.

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Macrophage Stimulation Protocols

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Macrophages were seeded either 1 day before stimulation or directly before stimulation into 96-well plates. Stimulation was done in 96-well plates for 6 h, and the medium was changed and further cultivated for additional 18 h. Immune cells were stimulated with rhIL-4 (2,000 U/mL, R&D Systems, #204-IL), rhIL-13 (200 U/mL, R&D Systems, #213-ILB), rhTNF-α (1,600 U/mL, R&D Systems, #210-TA), PGE₂ (2 µg/mL, Tocris, #2296), phorbol 12-myristate 13-acetate (PMA, 64 ng/mL, Sigma Aldrich, #P8139), rhIL-15 (200 U/mL, R&D Systems, #247-IL), or rhIFN-γ (200 U/mL, R&D Systems, #285-IF).

Sauer J., Steixner-Kumar A.A., Gabler S., Motyka M., Rippmann J.F., Brosa S., Boettner D., Schönberger T., Lempp C., Frodermann V., Simon E., Krenkel O, & Bahrami E. (2024). Diverse potential of secretome from natural killer cells and monocyte-derived macrophages in activating stellate cells. Frontiers in Immunology, 15, 1232070.

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Profiling RTK Signaling in Caco-2 Cells

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Confluent Caco-2 cells were fasted for 12h and then stimulated with rh-IFN-γ (10ng/ml) or rh-IL-13 (10ng/ml) (R&D Systems) for 5, 10, 45 and 90min. Cells were lysed and harvested as previously described in the presence of the protease inhibitor mixture and phosphatase inhibitor. The protein content of lysates was adjusted to 600µg/ml. The coated nitrocellulose membranes (PathScan RTK Signaling Antibody Array Kit, Cell Signaling Technology) were incubated with cell lysates over night at 4°C. The kinases captured by the antibody-coated spots were detected with a cocktail of biotinylated specific antibodies, then HRP-conjugated streptavidin and finally it was developed with Enhanced chemiluminescence reagents (ECL Plus; GE Healthcare, Danderyd, Sweden), following the manufacturer’s instructions. Membranes were scanned with the C digit scanner. The images were analyzed with the Image-J software.

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Inflammatory Immune Cell Analysis

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Recombinant human (rh)IL-4, rhIFN-γ and rhIL-13 were purchased from R&D Systems, Inc., (Minneapolis, MN, USA). Phorbol myristate acetate (PMA), BLM and LPS were supplied by Sigma-Aldrich; Merck KGaA (Darmstadt, Germany). BLM solutions were prepared with endotoxin-free saline to a final concentration of 10 U/ml and stored at −20°C. The anti-human-CD14-peridinin chlorophyll protein complex (PerCP; cat. no: 340585), CD68-fluorescein isothiocyanate (FITC; cat. no: 562117), CD80-allophycocyanin (APC; cat. no: 561134), CCR7-phycoerythrin (PE)-cyanine (cy)7(cat. no: 560922), CD206-PE(cat. no: 555954) and atched isotype antibodies were all supplied by BD Biosciences (Franklin Lakes, NJ, USA). FITC-dextran was purchased from Sigma-Aldrich; Merck KGaA.

Liu H., Dong H., Jiang L., Li Z, & Ma X. (2018). Bleomycin inhibits proliferation and induces apoptosis in TPC-1 cells through reversing M2-macrophages polarization. Oncology Letters, 16(3), 3858-3866.

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