Il 13

Manufactured by Thermo Fisher Scientific

Sourced in United States, United Kingdom, Germany, Austria, Canada

The IL-13 is a laboratory instrument designed for the detection and quantification of interleukin-13 (IL-13) levels in various biological samples. It utilizes advanced analytical techniques to provide accurate and reliable measurements of this important cytokine.

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Close spelling variants of this product

Recombinant human IL-13 (21 citations) IL-4/IL-13 (9 citations) Human IL-13 (8 citations) Recombinant murine IL-13 (6 citations) Interleukin 13 (IL-13) (4 citations) Recombinant human interleukin 13 (IL-13) (3 citations) IL-13 ELISA (3 citations) Human IL-13 ELISA kit (3 citations) IL-13-FITC (2 citations) Rat anti-mouse IL-13 (2 citations)

429 protocols using il 13

Quantification of Lung Cytokines by ELISA

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In order to determine lung tissue specific concentration of IL-4, IL-5, IL-13, IL-17, and IFN-γ, snap-frozen tissue pieces were homogenized in M-PER™ Mammalian Protein Extraction Reagent (ThermoFisherScientific), supplemented with PhosSTOP™ (Roche) and cOmplete™ protease Inhibitor cocktails (Roche). Protein concentration was determined with a Roti®-Quant protein quantification assay (Carl-Roth). For assessment of cytokine concentrations, 100 mg/ml total protein was used either IL-4 (eBioscience), IL-5 (Biolegend), IL-13 (eBioscience), IL-17 (Biolegend), or IFN-γ (eBioscience) specific ELISA assays. ELISA assays were carried out according to manufacturer's instructions.

Kindermann M., Knipfer L., Obermeyer S., Müller U., Alber G., Bogdan C., Schleicher U., Neurath M.F, & Wirtz S. (2020). Group 2 Innate Lymphoid Cells (ILC2) Suppress Beneficial Type 1 Immune Responses During Pulmonary Cryptococcosis. Frontiers in Immunology, 11, 209.

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In Vitro Model of Allergic Rhinitis

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hNECs procured from PromoCell GmbH were grown in RPMI‐1640 medium (Haoranbio) accompanied with the addition of 10% FBS (Biologic Industries) and 1% penicillin/streptomycin at 37°C under humidified air with 5% CO₂. Then, hNECs were administrated by IL‐13 (10 ng/ml; PeproTech, Inc.) for 24 h to mimic an in vitro model of AR,²⁵ regarding the non‐IL‐13 hNECs as a negative control group (Control). Additionally, hNECs were pretreated with SITR1 inhibitor EX527 (20 μM; Selleck Chemicals) for 24 h before IL‐13 exposure.²⁶

Xu H., Wang L., Chen H, & Cai H. (2022). HDAC4 depletion ameliorates IL‐13‐triggered inflammatory response and mucus production in nasal epithelial cells via activation of SIRT1/NF‐κB signaling. Immunity, Inflammation and Disease, 10(11), e692.

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Keratinocyte Hydrolysate Response to IL-13

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A pool of 4 x 10⁴ primary human epidermal keratinocytes (CELLnTec, Bern, Switzerland) were seeded in a 48-well plate (Corning, VWR, Nyon, Switzerland) in CnT57 medium (CELLnTec, Bern, Switzerland) supplemented with 10% serum. When keratinocytes reached 90% confluence, cells were treated with the hydrolysate at 1 µg protein/mL. One hour after the addition of the ingredient, 50 ng/mL of interleukin-13 (IL-13) (Peprotech, London, UK) was added and incubated at 37 °C with 5% CO₂ for 48 h. An identical volume of medium was added to the non-IL-13 primed cultures.

Holvoet S., Nutten S., Dupuis L., Donnicola D., Bourdeau T., Hughes-Formella B., Simon D., Simon H.U., Carvalho R.S., Spergel J.M., Koletzko S, & Blanchard C. (2021). Partially Hydrolysed Whey-Based Infant Formula Improves Skin Barrier Function. Nutrients, 13(9), 3113.

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Generating Primary and M2 Macrophages

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To obtain the primary human macrophages, PBMCs were treated with 100 ng/mL macrophage colony-stimulating factor (PeproTech) in RPMI (Roswell Park Memorial Institute) 1640 medium supplemented with 20% fetal bovine serum for 7 days. To generate M2 macrophages, the cells were treated with 20 ng/mL IL-4 (PeproTech) and 20 ng/mL IL-13 (PeproTech) for an additional 72 hours.
The polarization of M2 macrophages derived from THP-1 cells is described in our previous study.17 (link) Briefly, THP-1 cells were treated with 320 nM phorbol-12-myristate-13-acetate (PMA, Sigma) for 6 hours and then with PMA plus 20 ng/mL IL-4 (PeproTech) and 20 ng/mL IL-13 (PeproTech) for 18 hours to obtain the M2 phenotype.

Liu D., Xu X., Dai Y., Zhao X., Bao S., Ma W., Zha L., Liu S., Liu Y., Zheng J, & Shi M. (2021). Blockade of AIM2 inflammasome or α1-AR ameliorates IL-1β release and macrophage-mediated immunosuppression induced by CAR-T treatment. Journal for Immunotherapy of Cancer, 9(1), e001466.

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Macrophage Polarization and Cytokine Secretion

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The M1 phenotype was generated by 10 ng/mL LPS and 20 ng/mL interferon γ (IFN-γ; PeproTech), whereas M2 phenotype was generated by 20 ng/mL each IL-4 (PeproTech) and IL-13 (PeproTech) for 24 h. Next, IL-4- and IL-13-driven BMDM at a density of 3 × 10⁵/ml in 12-well plates were stimulated with 5 μg/mL LGp40, LGp40-C156S, CDp40, and CDp40-C155S at 37 °C for 48 h. Cultured supernatants were collected and the levels of IL-6, IL-12p40, and IL-10 were quantified using ELISA assay kits (R&D Systems). Lactate concentrations were determined using fluorometric quantification assay kits (Biovision).

Chen P.C., Hsieh M.H., Kuo W.S., Wu L.S., Kao H.F., Liu L.F., Liu Z.G., Jeng W.Y, & Wang J.Y. (2022). Moonlighting glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein of Lactobacillus gasseri attenuates allergic asthma via immunometabolic change in macrophages. Journal of Biomedical Science, 29, 75.

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Bone Marrow-Derived Macrophage Polarization

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Bone marrow cells were obtained by flushing the femurs and tibias of normal male C57/BL6 mice in Dulbecco's modified Eagle's medium (DMEM; GE Healthcare Life Sciences, Logan, UT, USA) supplemented with 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) and 30% L929 conditioned medium at 37°C. The medium was changed at day 3 and day 5. L929 conditioned medium was the supernatant from growing L929 cells in DMED-containing 10% FBS for 5 days. After 7 days incubation, bone marrow-derived macrophages (BMDMs, M0) were harvested. 100 ng/ml lipopolysaccharide (LPS) and 6 ng/ml interferon (IFN)-γ were used to stimulate M0 and M1 macrophages which were harvested after 24 h. In addition, M2 macrophages were induced by stimulating M0 macrophages with 10 ng/ml interleukin (IL)-4 and 10 ng/ml IL-13 for 24 h. IL-4, IL-13, LPS and IFN-γ were purchased from PeproTech, Inc. (Rocky Hill, NJ, USA).
Pioglitazone and PPARγ antagonist GW9662 were purchased from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany). M0 macrophages were stimulated with Pioglitazone (5 µm) for 24 h in the presence or absence of GW9662. PPARγ inhibitor GW9662 (5 µm) was added 6 h before treatment with Pioglitazone.

Zhang C., Zhang Y., Zhang C., Liu Y., Liu Y, & Xu G. (2019). Pioglitazone increases VEGFR3 expression and promotes activation of M2 macrophages via the peroxisome proliferator-activated receptor γ. Molecular Medicine Reports, 19(4), 2740-2748.

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NRVCM Proliferation Assay with STAT3 Inhibitor

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24 hours after plating, NRVCMs were serum starved (0% FBS) in DMEM for 3 hours. Following serum starvation cells were treated in triplicate with 10μM BrdU and either 0nM, 100nM, or 1μm of STAT3 inhibitor (Stattic, Santa Cruz sc-202818) in serum free (0% FBS) DMEM media. IL13 treated wells also received 20ng/mL IL13 (Peprotech). After 18 hours of incubation in 37°C, treated NRVCMs were fixed in 4% PFA for 10 min and then washed in PBS. Fixed cells were permeabilized with 4N HCl and 0.5% Triton X-100 in PBS for 10 min, blocked with 2% GS in PBS, and stained with primary antibodies diluted 1:400 in 2% GS for 2 hours. Cells were washed with PBS, and then incubated for 30 min in secondary antibodies diluted 1:400 in 2% GS, followed by DAPI incubation for 10 min. Cells were imaged with a 20X objective on PCO.panda 4.2M camera on Nikon Eclipse 80i microscope.

Paddock S.J., Swift S.K., Alencar-Almeida V., Kenarsary A., Alvarez-Argote S., Flinn M.A., Patterson M, & O’Meara C.C. (2021). IL4Rα Signaling Promotes Neonatal Cardiac Regeneration and Cardiomyocyte Cell Cycle Activity. Journal of molecular and cellular cardiology, 161, 62-74.

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Cytokine Quantification in Biological Samples

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Supernatants collected from re-stimulated lymph node cells or BAL samples were added to Nunc Maxisorp 96-well plates (Thermo Fisher Scientific), which had been coated with purified anti-mouse IL-4 antibody (Thermo Fisher Scientific), IL-5 (BD Biosciences), IL-13 (Thermo Fisher Scientific) and IFN-γ (Thermo Fisher Scientific) all at 2 μg/ml O/N at 4 °C. Plates were subsequently blocked with 1% BSA. Cytokines were detected with biotinylated anti-mouse IL-4 (Thermo Fisher Scientific), IL-5 (Thermo Fisher Scientific), IL-13 (Thermo Fisher Scientific) and IFN-γ (Thermo Fisher Scientific) all at 0.5 μg/ml. Recombinant IL-4, IL-5, IL-13 and IFN-γ (R&D systems) were used as a standard (starting concentration 10 ng/ml). Antibody binding was detected with streptavidin-HRP (BD Biosciences) and developed with TMB substrate set (BD Biosciences).

Agaronyan K., Sharma L., Vaidyanathan B., Glenn K., Yu S., Annicelli C., Wiggen T.D., Penningroth M.R., Hunter R.C., Dela C.S, & Medzhitov R. (2022). Tissue remodeling by an opportunistic pathogen triggers allergic inflammation. Immunity, 55(5), 895-911.e10.

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Macrophage Polarization Protocol

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M0-Mφs were polarized into M1-Mφs by treatment with 20 ng/ml of IFN-γ (Invitrogen, Thermo Fisher, Waltham, Massachusetts, USA) and 100 ng/ml of LPS (InvivoGen, Toulouse, France) for 24 or 48 h. On the other hand, M0-Mφs were polarized into M2-Mφs by incubation with 20 ng/ml of IL-13 (Gibco, Thermo Fisher, Waltham, Massachusetts, USA) and 20 ng/ml of IL-4 (Invitrogen) for 24 or 48 h.

Bazzi S., Frangie C., Azar E, & Daher J. (2022). The effect of myeloperoxidase-oxidized LDL on THP-1 macrophage polarization and repolarization. Innate Immunity, 28(2), 91-103.

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Murine Macrophage Polarization Protocol

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RAW 264.7 murine macrophages (ATCC TIB-71) were maintained in culture medium composed from standard RPMI 1640 (Gibco), 10% fetal bovine serum, and 1% penicillin:streptomycin. RAW macrophages were seeded in 35 mm glass bottom dishes (MatTek) for imaging experiments. All imaging samples were plated at a density of 1×10⁵ cells per dish and incubated at 37 ⁰C and 5% CO₂, for 24 hours to allow cell adhesion. Separate cultures of macrophages polarized with M1-like (IFN-γ) or M2-like (IL-4/IL-13) cytokines were generated by standard media substitution with 2 mL cytokine-supplemented media and incubated between 24–72 hours. Media for M(IFN-γ) stimulation consisted of RPMI 1640 supplemented with 10 ng/ml interferon-γ (IFN-γ; R&D systems), and media for M(IL4/IL13) stimulation consisted of RPMI 1640 with 20 ng/ml interleukin 4 (IL-4; Invitrogen) and 20 ng/ml interleukin 13 (IL-13; Gibco).

Heaster T.M., Humayun M., Yu J., Beebe D.J, & Skala M.C. (2020). Autofluorescence imaging of 3D tumor-macrophage microscale cultures resolves spatial and temporal dynamics of macrophage metabolism. Cancer research, 80(23), 5408-5423.

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