Il 17

Manufactured by Thermo Fisher Scientific

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

The IL-17 is a lab equipment product designed for the detection and quantification of the interleukin-17 (IL-17) cytokine. IL-17 is a pro-inflammatory cytokine involved in various immune responses. The IL-17 product provides a reliable and accurate method for researchers to measure IL-17 levels in biological samples.

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Lab products found in correlation

Close spelling variants of this product

Anti-IL-17 (ebio17B7) IL‐17 human ELISA Kit PE-conjugated anti-IL-17 mAb Recombinant human IL-17 PE-conjugated anti-IL-17 IL-17-PerCP-Cy5.5 IL-17-FITC Anti-CD161-PE (clone HP-3G10) and anti-IL-17-Alexa488 (clone eBio64DEC17) Anti-IL-17 IL-17-PE

182 protocols using il 17

Shikonin Modulates IL-17-Induced HaCaT Cell Proliferation

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The proliferation of HaCaT cells was assessed using a MTS assay (Promega Corporation) according to the manufacturer's instructions. Briefly, the cells were seeded into 96-well plates, at a density of 5,000 cells/well and cultured for 24, 48 and 72 h in the presence of varying concentrations of shikonin (0.00, 0.01, 0.05, 0.25, 0.50, 1.00, 2.50 or 5.00 µM) (Sigma-Aldrich; Merck KGaA) in a humidified incubator at 37°C with 5% CO₂. In order to verify the effect of shikonin on IL-17 induced HaCaT cell proliferation, the HaCaT cells were seeded into 96-well plates, at a density of 5,000 cells/well and cultured in a humidified incubator at 37°C for 24, 48 and 72 h with IL-17 (PeproTech, Inc.), shikonin, IL-17 + shikonin, or no treatment. In order to verify the effect of shikonin on IL-17-induced HaCaT cell proliferation when CEBPD is silenced, the assay was performed with scrambled siRNA (NC), RNAi + IL-17, RNAi + shikonin, RNAi + IL-17 + shikonin, and untreated cells. The number of cells and the duration of treatment are consistent with the previous description. The absorbance was measured in each well at 490 nm using a microplate reader (Bio-Rad Laboratories, Inc.). Each sample was analyzed in six replicates, and the assay was repeated three times.

Lan X.O., Wang H.X., Qi R.Q., Xu Y.Y., Yu Y.J., Yang Y., Guo H., Gao X.H, & Geng L. (2020). Shikonin inhibits CEBPD downregulation in IL-17-treated HaCaT cells and in an imiquimod-induced psoriasis model. Molecular Medicine Reports, 22(3), 2263-2272.

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Cytokine Quantification by Sandwich ELISA

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The concentrations of IL-4, IL-5, IL-10, IL-13, IL-17, and IFN-γ in supernatants were quantified by sandwich ELISA using the following anticytokine antibody pairs: IL-4 (Biosource; AHC0642 and AHC0749), IL-5 (BioLegend; 500902 and 501002), IL-10 (eBioscience; 14-7108-85 and 13-7109-85), IL-13 (Invitrogen; P130E and M130B), IL-17 (eBioscience; 14-7178-85 and 13-7179-85), and IFN-γ (eBioscience; 14-7318-85 and 13-7319-85). The capture antibody was coated in NUNC MaxiSorp plates (Greiner Bio-One) overnight at 4°C in 0.05 mol Na₂CO₃ buffer (pH 9.7) (Merck, Boston, Mass). The plates were blocked with 1% BSA for 1 hour at 37°C. Supernatants and biotinylated antibodies were added and plates were incubated for 2 hours at room temperature.

Keumatio Doungtsop B.C., Nardini E., Kalay H., Versteeg S.A., Lübbers J., van Barneveld G., Li E.R., van Vliet S.J., van Ree R., de Jong E.C, & van Kooyk Y. (2023). Sialic acid–modified der p 2 allergen exerts immunomodulatory effects on human PBMCs. The Journal of Allergy and Clinical Immunology: Global, 3(1), 100193.

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Osteoclast Differentiation Modulation

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RAW264.7 (4.5 × 10⁴) cells were seeded into 48-well plate and differentiated into osteoclasts in the presence of the receptor activator of NF-κB ligand (RANKL, 75 ng/mL; Peprotech) with or without proinflammatory cytokines (IL-12, IL-23, IFN-γ, and IL-17 alone: 75 ng/mL; IL-12 + IL-23: 37.5 ng/mL each; IL-12 + IL-23 + IFN-γ + IL-17: 18.75 ng/mL each; Peprotech). The medium was changed every 2 days. After 3 days of osteoclast differentiation, the cells were fixed with 4% PFA and stained with a TRAP kit (Sigma-Aldrich).

Xu J., Li J., Hu Y., Dai K., Gan Y., Zhao J., Huang M, & Zhang X. (2020). IL-23, but not IL-12, plays a critical role in inflammation-mediated bone disorders. Theranostics, 10(9), 3925-3938.

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Cytokine Production Profiling of T-Cell Clones

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To assess the cytokines production, 10⁶ T-cell blasts of each clone were stimulated in duplicate cultures after stimulation for 36 hours with anti-CD3 monoclonal antibody or with PMA plus anti-CD3monoclonal antibody. Cell-free supernatants were collected and assayed for their cytokine content (IFN-γ, IL-4, IL-17, and tumor necrosis factor-α; Bio-Source International, Camarillo, CA) as previously described.^{[21 (link)]} Supernatants showing IFN-γ, IL-4, IL-17, and TNF-α levels 5 SD over the mean levels in control supernatants derived from irradiated feeder cells alone were regarded as positive. T-cell clones able to produce IFN-γ but not IL-4, nor IL-17 were categorized as Th1; clones able to produce IL-4 but not IFN-γ, nor IL-17 as Th2; clones producing IL-17, but not IFN-γ nor IL-4 as Th17; clones producing both IFN-γ and IL-17, but not IL-4 as Th17/Th1; and clones producing both IFN-γ and IL-4, but not IL-17 as Th0, as previously described.^{[22 (link)]}

Emmi G., Silvestri E., Bella C.D., Grassi A., Benagiano M., Cianchi F., Squatrito D., Cantarini L., Emmi L., Selmi C., Prisco D, & D’Elios M.M. (2016). Cytotoxic Th1 and Th17 cells infiltrate the intestinal mucosa of Behcet patients and exhibit high levels of TNF-α in early phases of the disease. Medicine, 95(49), e5516.

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Multiplex Cytokine Profiling in Plasma

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In vitro samples and plasma samples were analysed in Leiden, The Netherlands and Copenhagen, Denmark, respectively, both on the Luminex platform (Luminex 100, Luminex Corp., Austin, TX, USA). The in vitro stimulation assay was slightly modified from a previous study [9] , with the following analytes (lower limit of detection (LLD) given in parenthesis): TNF-α (10 pg/ml), IFN-γ (5 pg/ml), IL-5 (3 pg/ml), IL-10, IL-13 (10 pg/ml) and IL-17 (10 pg/ml) (BioSource, Camarillo, CA, USA). In plasma, TNF-α (5 pg/ml), IL-10 (5 pg/ml), IL-6 (7 pg/ml), MCP-1 (10 pg/ml), IL-8 (2.8 pg/ml), IL-1Ra (30 pg/ml) were measured (Fluorokine MAP Multiplex Human Cytokine Panel A, R&D Systems, Minneapolis, MN, USA). Concentrations of suPAR (0.1 ng/ml), a marker of inflammation [10] (link), were measured from plasma by enzyme-linked immunosorbent assay (ELISA) (suPARnostic, ViroGates, Birkerød, Denmark). Measurements below the LLD were referred to as non-detectable (ND). Baseline and follow-up samples from each child were measured on the same assay plate.

Jensen K.J., Søndergaard M., Andersen A., Sartono E., Martins C., Garly M.L., Eugen-Olsen J., Ullum H., Yazdanbakhsh M., Aaby P., Benn C.S, & Erikstrup C. (2014). A Randomized Trial of an Early Measles Vaccine at 4½ Months of Age in Guinea-Bissau: Sex-Differential Immunological Effects. PLoS ONE, 9(5), e97536.

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Cytokine Production by IF-specific T Cells

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To assess the cytokine production of IF-specific T cell clones upon antigen stimulation, 5 × 10⁵ T cell blasts of each clone were co-cultured for 48 h in 0.5 ml of serum-free medium with 5 × 10⁵ irradiated autologous PBMCs in the absence or presence of IF [13 (link)]. At the end of the culture period, duplicate samples of each supernatant were assayed for their IFN-γ, TNF-α, IL-4, IL-21 and IL-17 (BioSource International, Camarillo, CA) production by ELISA. For further investigation, T cell blasts from each IF-specific T cell clone were stimulated with medium or IF (10 μg/ml) in the presence of autologous APCs for 48 h in ELISPOT microplates coated with anti- IFN-γ or anti-IL-17 antibody, respectively (eBioscience, Inc., San Diego, Ca, USA). At the end of culture period, the number of IFN-γ and IL-17 SFCs were counted as described [21 (link)].

Troilo A., Grassi A., Petrone L., Cianchi F., Benagiano M., Bella C.D., Capitani N., Bitetti J., D’Elios S., Tapinassi S., Azzurri A., Alnwaisri H., Romagnoli J., Bizzaro N., Bergman M., Baldari C.T, & D’Elios M.M. (2019). Intrinsic factor recognition promotes T helper 17/T helper 1 autoimmune gastric inflammation in patients with pernicious anemia. Oncotarget, 10(30), 2921-2929.

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Cytokine Profiling of Activated T Cells

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Single cell suspensions from spleen and axillary lymph nodes were prepared as described above. The cell viability, generally around 90%, was analyzed using Trypan Blue (Sigma-Aldrich) exclusion and cells were seeded at 2 x 10⁵ of live cells per well in 96-well plates in RPMI 1640 (Sigma-Aldrich) culture medium supplemented with 10% fetal bovine serum (BioClot GmbH, Aidenbach, Germany) and 1% Antibiotic-Antimycotic solution (Sigma-Aldrich). Then, we stimulated the cells for 48 hours with plate-bound anti-CD3 (5 μg/ml; clone 145-2C11) and soluble anti-CD28 (2 μg/ml; clone 37.51; both eBioscience) antibodies. The supernatants were collected and frozen at -20°C until analysis. Commercially available ELISA sets were used to measure the levels of IFN-γ and IL-17 (Invitrogen Corp.) in the supernatants. All tests were performed according to the manufacturers' recommendations.

Zákostelská Z., Málková J., Klimešová K., Rossmann P., Hornová M., Novosádová I., Stehlíková Z., Kostovčík M., Hudcovic T., Štepánková R., Jůzlová K., Hercogová J., Tlaskalová-Hogenová H, & Kverka M. (2016). Intestinal Microbiota Promotes Psoriasis-Like Skin Inflammation by Enhancing Th17 Response. PLoS ONE, 11(7), e0159539.

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Murine Cytokine and Immune Markers Quantification

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Murine interleukin (IL)-6, IL-8, tumor necrosis factor (TNF)-α, IL-17, IL-18 and IL-1β enzyme-linked immunosorbent assay (ELISA) kits were purchased from Invitrogen Life Technologies; Thermo Fisher Scientific, Inc. (Waltham, MA, USA). Rat anti-mouse-Ly-6 G (cat. no. 551495) monoclonal antibodies (mAbs) were purchased from BD Pharmingen (San Diego, CA, USA). Rat anti-mouse F4/80 (clone A3-1) mAb (cat. no. MCA497GA) was acquired from Serotec (Bio-Rad Laboratories, Inc., Hercules, CA, USA). FITC-conjugated sheep anti-rat IgG mAb (cat. no. PA1-28638) was purchased from Pierce (Thermo Fisher Scientific, Inc.). Rabbit anti mouse SP-C polyclonal Abs (cat. no. bs-10067R) was acquired from Bioss Inc. (Woburn, MA, USA). Total protein, albumin and keratinocyte growth factor (KGF) ELISA kits and Texas Red-conjugated goat anti-rabbit IgG (H + L) (cat. no. ABIN287315) were acquired from Antibodies-Online GmbH (Aachen, Germany).

Ding Q., Liu G., Zeng Y., Zhu J., Liu Z., Jiang J, & Huang J. (2017). Glycogen synthase kinase-3β inhibitor reduces LPS-induced acute lung injury in mice. Molecular Medicine Reports, 16(5), 6715-6721.

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Gastric Carcinoma Cell Line Responses

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The human gastric carcinoma cell line, HGC-27, was obtained from the American Type Culture Collection. Cells were cultured in Dulbecco's modified Eagle's medium (DMEM, Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (HyClone; Cytiva), 100 U/ml penicillin and 100 U/ml streptomycin (Invitrogen; Thermo Fisher Scientific, Inc.) in an incubator with 5% CO₂ at 37˚C. Cell propagation was performed every 24 h. The cells in the logarithmic phase were used for further analysis.
After cells were fully adhered to the well, they were cultured at 37˚C at a density of 2x10³ cells/cm² and divided into the following groups: i) Control group; ii) IL-17 group; iii) IL-17-apatinib group; and iv) apatinib group. Cells in the IL-17 group were treated with 10 ng/ml IL-17 (Invitrogen; Thermo Fisher Scientific, Inc.) for 48 h. The apatinib group was treated with 50 ng/ml apatinib (cat. no. S7297, Selleck Chemicals) for 48 h. For the IL-17-apatinib group, cells were cultured with 10 ng/ml IL-17 and 50 ng/ml apatinib for 48 h. Cells in the Control group were treated with same volume of DMEM. All treatments were performed at 37˚C.

Wang T., Zhang J, & Cui L. (2021). Apatinib inhibits gastric carcinoma development by regulating the expression levels of IL-17 via the Bax/Bcl-2 signaling pathway. Experimental and Therapeutic Medicine, 21(6), 654.

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Overexpression and Knockdown of MMP7 in Prostate Cancer Cells

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Human prostate cancer cell lines (LNCaP, C4-2B, and PC-3) were authentic and free of mycoplasma as described previously⁵⁹ and cultured in monolayer in a humidified 5% CO₂ incubator at 37°C. The cell lines were transfected with human MMP7 plasmid (Addgene, Cambridge, MA, USA) using Lipofectamine^® 2000 Transfection Reagent (Invitrogen, Grand Island, NY, USA) according to the manufacturer’s instructions. Positive cells were selected with 1 mg/ml Geneticin^® (Invitrogen) for two weeks and the cells stably overexpressing MMP7 were named LNCaP-MMP7, C4-2B-MMP7, PC-3-MMP7, respectively. The cells were treated without or with 20 ng/ml recombinant human IL-17 (R&D Systems) for 0.5, 2, and 8 hours. In another experiment, the cells were first transfected with 5 nM control siRNA (catalog #4390843, Silencer^® Select Negative Control) or 5 nM siRNA targeting MMP7 (catalog #4392420, Silencer^® Select Pre-designed siRNA) (Fisher Scientific) using Lipofectamine^® 2000 Transfection Reagent (Invitrogen) according to the manufacturer’s instructions; 24 hours later, the cells were treated without or with IL-17 for 2 and 8 hours.

Zhang Q., Liu S., Parajuli K., Zhang W., Zhang K., Mo Z., Liu J., Chen Z., Yang S., Wang A., Myers L, & You Z. (2016). Interleukin-17 promotes prostate cancer via MMP7-induced epithelial-to-mesenchymal transition. Oncogene, 36(5), 687-699.

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