Recombinant human il 13

Manufactured by Thermo Fisher Scientific

Sourced in United States

Recombinant human IL-13 is a cytokine that is produced in E. coli. It has a molecular weight of approximately 12 kDa.

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Lab products found in correlation

Recombinant human il 4, by Thermo Fisher Scientific (5 mentions) Recombinant human m csf, by Thermo Fisher Scientific (2 mentions) Facs celesta sorp flow cytometer, by BD (2 mentions) Ifn γ, by Thermo Fisher Scientific (2 mentions) Lps e055 b55, by Merck Group (2 mentions) Legendplex human macrophage microglia panel, by BioLegend (2 mentions) Inverted bright field microscope, by Leica (1 mentions) 24 well plate, by Thermo Fisher Scientific (1 mentions) Dulbecco modified eagle medium (dmem), by Merck Group (1 mentions) Ampicillin, by Merck Group (1 mentions) Dexamethasone (dex), by Fujifilm (1 mentions) Cell count reagent sf, by Nacalai Tesque (1 mentions) Clarithromycin, by Fujifilm (1 mentions) Erythromycin, by Fujifilm (1 mentions) Josamycin, by Merck Group (1 mentions) Ampicillin, by Fujifilm (1 mentions) Anti rabbit igg hrp linked antibody, by Cell Signaling Technology (1 mentions) Alexa fluor 488 dye, by Thermo Fisher Scientific (1 mentions) Normal mouse igg, by Santa Cruz Biotechnology (1 mentions) Mouse monoclonal β actin antibody, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

IL-13 (429 citations) Human IL-13 (8 citations) Recombinant IL-13 (3 citations) Recombinant human interleukin 13 (IL-13) (3 citations) Recombinant human IL-4 and IL-13 (3 citations) Recombinant human TNF‐α and IL‐13 (2 citations) IDM and/or recombinant human IL-13 (2 citations)

21 protocols using recombinant human il 13

Polarized Bronchial Epithelial Cell Culture

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Polarized human bronchial epithelial cell cultures (MucilAir) were provided by Epithelix (Sàrl Epithelix). The primary cells were generated from a single healthy donor (71-year-old female with no smoking history). The MucilAir cell culture medium was changed every 2 days until experimentation. Respective wells were treated with MucilAir cell culture medium containing AcPGP (10 μg/ml; AnaSpec peptides), human recombinant IL-13 (10 ng/ml; Peprotech), or a combination of AcPGP (10 μg/ml) and human recombinant IL-13 (10 ng/ml). MucilAir cell culture medium was used for media control wells. The respective wells were treated apically overnight and then continuously fed basolaterally every 2 days with medium containing respective treatments. Apical supernatant was collected by washing the apical side of the Transwell with 200 μl of PBS on day 7 and stored at −80°C for subsequent analysis. At the end of the experiment, inserts were fixed in 4% paraformaldehyde and processed for histological analysis.

Patel D.F., Peiró T., Shoemark A., Akthar S., Walker S.A., Grabiec A.M., Jackson P.L., Hussell T., Gaggar A., Xu X., Trevor J.L., Li J., Steele C., Tavernier G., Blalock J.E., Niven R.M., Gregory L.G., Simpson A., Lloyd C.M, & Snelgrove R.J. (2018). An extracellular matrix fragment drives epithelial remodeling and airway hyperresponsiveness. Science translational medicine, 10(455), eaaq0693.

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SARS-CoV-2 Spike Protein Modulates Macrophage Polarization

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To polarize human macrophages, monocyte derived macrophages were plated in 48 well plates at 5 × 10⁴ cells per well in 500μl RPMI medium and allowed to rest 2hours at 37°C before treat of cytokines or LPS. The human M0 macrophages were either left untreated or treated for 48h to induce macrophage polarization: (i) for M1 polarization with 10 ng/mL LPS (E055:B55; Sigma-Aldrich) and 20 ng/mL IFNγ (PeproTech), (ii) for M2 polarization with 50 ng/ml human recombinant M-CSF, 20 ng/ml human recombinant IL-4, and 20 ng/ml human recombinant IL-13 (PeproTech). Human macrophages were then cultured alone or with 0.1ug, or 1ug SARS-CoV-2 Spike proteins for 48 hr. The cultured supernatants were collected, and cytokine levels determined using the bead-based immunoassays LEGENDplex^™ Human Macrophage/ Microglia Panel (Biolegend). The assays were performed in 96-well plates following the manufacturer’s instructions. For measurements a FACSCelesta SORP flow cytometer (BD Biosciences) was employed, and data were evaluated with the LEGENDplex^™ Data Analysis software.

Park C., Hwang I.Y., Yan S.L., Vimonpatranon S., Wei D., Van Ryk D., Girard A., Cicala C., Arthos J, & Kehrl J.H. (2023). Murine Alveolar Macrophages Rapidly Accumulate Intranasally Administered SARS-CoV-2 Spike Protein leading to Neutrophil Recruitment and Damage. bioRxiv.

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Colony Formation Assay for CML Cells

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K562 cell line (1,000 cells/well) or CD34⁺ CML sorted cells (10,000 cells/well) were seeded in 6-well plates in 1.5 mL MethoCult Matrix (H4100, StemCell) after treatment for 48 hours with human recombinant IL-13 (50 ng/mL, Peprotech) or medium only. Cells were cultured for 12-14 days to allow the colonies to form. Formed colonies were then scored after incubation at 37°C in a fully humidified 5% CO₂ atmosphere. Counting was performed manually by using an inverted brightfield microscope (Leica) at 10x magnification.

Fiordi B., Salvestrini V., Gugliotta G., Castagnetti F., Curti A., Speiser D.E., Marcenaro E., Jandus C, & Trabanelli S. (2023). IL-18 and VEGF-A trigger type 2 innate lymphoid cell accumulation and pro-tumoral function in chronic myeloid leukemia. Haematologica, 108(9), 2396-2409.

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Polarizing Macrophages for SARS-CoV-2 Spike Protein Interactions

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To polarize human macrophages, monocyte-derived macrophages were plated in 48-well plates at 5×10⁴ cells per well in 500 µl RPMI medium and allowed to rest for 2 hr at 37°C before treating cytokines or LPS. The human M0 macrophages were either left untreated or treated for 48 hr to induce macrophage polarization: (i) for M1 polarization with 10 ng/ml LPS (E055:B55; Sigma-Aldrich) and 20 ng/ml IFNγ (PeproTech), (ii) for M2 polarization with 50 ng/ml human recombinant M-CSF, 20 ng/ml human recombinant IL-4, and 20 ng/ml human recombinant IL-13 (PeproTech). Human macrophages were then cultured alone or with 0.1 or 1 µg SARS-CoV-2 Spike proteins for 48 hr. The cultured supernatants were collected, and cytokine levels determined using the bead-based immunoassays LEGENDplex Human Macrophage/Microglia Panel (BioLegend). The assays were performed in 96-well plates following the manufacturer’s instructions. For measurements, a FACSCelesta SORP flow cytometer (BD Biosciences) was employed, and data were evaluated with the LEGENDplex Data Analysis software.

Park C., Hwang I.Y., Yan S.L., Vimonpatranon S., Wei D., Van Ryk D., Girard A., Cicala C., Arthos J, & Kehrl J.H. (2024). Murine alveolar macrophages rapidly accumulate intranasally administered SARS-CoV-2 Spike protein leading to neutrophil recruitment and damage. eLife, 12, RP86764.

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Antibiotic and Dexamethasone Effects on IL-13-stimulated MRC5 Cells

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MRC5 cells, a human embryonic lung fibroblast cell line (Riken BioResource Center, Tsukuba, Japan), were cultured with Dulbecco modified Eagle medium (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% fetal calf serum, 100 μg/mL streptomycin, and 100 U/mL penicillin G. MRC5 cells (7 × 10⁴ cells per well) were placed in 24-well plates (Nunc, Roskilde, Denmark) and cultured in 5% CO₂ humidified atmosphere at 37 °C with or without clarithromycin, erythromycin (Wako Pure Chemical Industries, Osaka, Japan), josamycin (Sigma-Aldrich), ampicillin (Sigma-Aldrich), or dexamethasone (Wako Pure Chemical Industries). clarithromycin was kindly supplied by Taisho Toyama Co., Ltd. (Tokyo, Japan). clarithromycin, erythromycin, josamycin, and ampicillin were dissolved in ethanol (EtOH, Wako) to therapeutic concentrations [19 (link), 20 (link)]. dexamethasone was dissolved in EtOH to 100 nM [21 (link)]. The final concentration of EtOH added to cells was 0.5%. After 24 h of culture, cells were stimulated by 50 ng/mL human recombinant IL-13 (Peprotech, Rocky Hill, NJ, USA) for 24 h. Cell viability was evaluated using WST-8 assay (Cell Count Reagent SF, Nacalai Tesque, Kyoto, Japan).

Komiya K., Ohta S., Arima K., Ogawa M., Suzuki S., Mitamura Y., Nunomura S., Nanri Y., Yoshihara T., Kawaguchi A., Kadota J.I., Rubin B.K, & Izuhara K. (2017). Clarithromycin attenuates IL-13–induced periostin production in human lung fibroblasts. Respiratory Research, 18, 37.

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Effects of Cigarette Smoke on Airway Cells

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The effects of CS on OPN production in ALI-PBECs differentiated in the presence of IL-13, which promotes goblet cell hyperplasia, were investigated in this experimental model. PBECs were grown to confluence and cultured at the ALI for 14 days in the presence and absence of recombinant human IL-13 (2.5 ng/mL, Peprotech, Rocky Hill, NJ, USA), which was added to the basolateral compartment of Transwell inserts. This was followed by exposure to whole CS or air once daily for a further 5 days in the presence and absence of continued IL-13 treatment (2.5 ng/mL), which was added to the basolateral compartment of the wells. Medium was refreshed every 2 days during the first 14 days and daily during the last 5 days, directly after whole CS or air exposure. Medium from the basolateral compartment and cells from the apical compartment were collected 24 h after final exposure to whole CS or air^{25 (link)}.

Ali M.N., Mori M., Mertens T.C., Siddhuraj P., Erjefält J.S., Önnerfjord P., Hiemstra P.S, & Egesten A. (2019). Osteopontin Expression in Small Airway Epithelium in Copd is Dependent on Differentiation and Confined to Subsets of Cells. Scientific Reports, 9, 15566.

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Differentiation of THP1 Cells into TAM Model

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The in vitro TAM model was generated as follows. Briefly, THP1 cells (5 × 10⁵ cells/mL) were differentiated using 200 nM phorbol 12-myristate 13-acetate (PMA) (32 (link)) (Solarbio, Cat. no.: p6741) for different time-periods (0, 24, 48, 72, and 96 h); we observed the changes in cellular morphology under an inverted phase-contrast microscope. Differentiation of PMA-treated cells was enhanced after the initial 3-d impetus by removing the PMA-containing medium and then incubating the cells in fresh RPMI 1640 with 20 ng/mL recombinant human IL-4 (PeproTech, Cat. no.: 200-04] and 20 ng/mL recombinant human IL-13 (PeproTech, Cat. no.: 200-13) for different time-periods (0, 24, 48, and 72 h) The complete medium was then replaced and cultured for another 2 days, and the supernatant was collected for subsequent experiments.

Le F., Yang L., Han Y., Zhong Y., Zhan F., Feng Y., Hu H., Chen T, & Tan B. (2021). TPL Inhibits the Invasion and Migration of Drug-Resistant Ovarian Cancer by Targeting the PI3K/AKT/NF-κB-Signaling Pathway to Inhibit the Polarization of M2 TAMs. Frontiers in Oncology, 11, 704001.

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Recombinant IL-13 Signaling Pathway Modulation

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Recombinant human IL-13 (Peprotech, Rocky Hill, NJ, USA) was dissolved in distilled water and added to the culture medium at a final concentration of 1, 5, or 10 ng/mL. The antibodies for immunofluorescence and immunohistochemistry staining were used as follows: Anti-IL-13Rα2 mouse monoclonal antibody (Abcam, Cambridge, UK), normal mouse IgG (Santa Cruz Biotechnology, Dallas, TX, USA), and goat anti-mouse IgG conjugated with Alexa Fluor 488 dye (Thermo Fisher Scientific, Waltham, MA, USA). The antibodies for western blotting were used as follows: Anti-ERK1/2, JNK, p38 MAPK, phospho-ERK1/2 (Thr202/Tyr204), phospho-JNK (Thr183/Tyr185), and phospho-p38 MAPK (Thr180/Tyr182) rabbit monoclonal antibodies and β-actin mouse monoclonal antibody (Cell Signaling Technology, Danvers, MA, USA) as primary antibodies, and anti-mouse IgG and anti-rabbit IgG HRP-linked antibody (Cell Signaling Technology) as secondary antibodies. Signal transduction inhibitor U0126 (ERK1/2 inhibitor) was purchased from Cell Signaling Technology. SP600125 (JNK inhibitor) and SB203580 (p38 inhibitor) were obtained from Tocris Bioscience (Bristol, UK).

Ulzii D., Kido-Nakahara M., Nakahara T., Tsuji G., Furue K., Hashimoto-Hachiya A, & Furue M. (2019). Scratching Counteracts IL-13 Signaling by Upregulating the Decoy Receptor IL-13Rα2 in Keratinocytes. International Journal of Molecular Sciences, 20(13), 3324.

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Cardiac Fibroblast Polarization Assay

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We acquired human primary cardiac fibroblasts from Cell Applications, Inc (San Diego, CA). Cells were thawed at 37°C immediately after taken out from liquid nitrogen (−196°C), washed and seeded on uncoated culture flasks in human cardiac fibrob-lasts growth medium according to manufacturer’s instructions (Cell Applications). Cells were passaged every 2–3 days. For stimulation, cells were seeded on 6-well plates and stimulated with 50 ng/mL of recombinant human IL-17A (Peprotech) for the Th17 conditions or 100 ng/mL of recombinant human IL-13 (Peprotech) for the Th2 conditions for three days. For flow cytometry, cells were detached using 0.05% trypsin-EDTA (Quality Biologicals) and washed in PBS prior to antibody staining. For cytokine repolarization experiments, cell culture supernatant was harvested, and cells were washed with PBS and cultured for another three days with new stimuli according to experiment design. Unstimulated cells were used as controls. Supernatant was collected upon harvest and stored at −80°C. Secreted cytokine levels were measured by quantitative sandwich ELISA.

Chen G., Bracamonte-Baran W., Diny N.L., Hou X., Talor M.V., Fu K., Liu Y., Davogustto G., Vasquez H., Taegtmeyer H., Frazier O.H., Waisman A., Conway S.J., Wan F, & Čiháková D. (2018). Sca-1+ cardiac fibroblasts promote development of heart failure. European journal of immunology, 48(9), 1522-1538.

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Airway Epithelial Cell Culture with IL-13

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Following the establishment of ALI, cells were fed basolaterally on alternate days with ALI medium [5] supplemented with recombinant human IL-13 (PeProTech EC Ltd, UK) at 20 ng/ml throughout the duration of the cultures (28 days) starting at day 0 ALI [11] , [23] (link)–[30] (link).

Thavagnanam S., Parker J.C., McBrien M.E., Skibinski G., Shields M.D, & Heaney L.G. (2014). Nasal Epithelial Cells Can Act as a Physiological Surrogate for Paediatric Asthma Studies. PLoS ONE, 9(1), e85802.

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