Human bronchial SMCs from ScienCell Research Laboratories (Carlsbad, CA, USA) were cultured in accordance with the manufacturer’s instructions in 5% CO2 at 37°C. Cells in passages 3–7 were used. Cells were growth-arrested for 2 days in media containing 0.01% fetal bovine serum for proliferation assays or serum-starved overnight for signaling studies. Cells were treated with recombinant human IL-22 (PeproTech, Inc., Rocky Hill, NJ, USA) or human recombinant PDGF-BB (Invitrogen, Carlsbad, CA, USA). For siRNA knockdown, cells were transfected with an siRNA Transfection Reagent and gene silencing siRNAs along with the controls by using a scrambled sequence from Santa Cruz Biotechnology (Dallas, TX, USA). Cells were used for experiments 2 days after transfection.
Recombinant human il 22
Manufactured by Thermo Fisher Scientific
Sourced in United States
Recombinant human IL-22 is a cytokine protein produced in a laboratory setting. It is a member of the IL-10 cytokine family and plays a role in immune system regulation and tissue repair processes.
Automatically generated - may contain errors
Lab products found in correlation
Recombinant human tnf α, by Thermo Fisher Scientific (2 mentions)
Sirna transfection reagent, by Santa Cruz Biotechnology (1 mentions)
Recombinant human il 17a, by Thermo Fisher Scientific (1 mentions)
Antibody against gapdh, by Proteintech (1 mentions)
Pe annexin 5 apoptosis detection kit 1, by BD (1 mentions)
Wst 1 cell proliferation reagent, by Takara Bio (1 mentions)
Dmem 1 g l glucose, by Thermo Fisher Scientific (1 mentions)
Primary human hepatocytes, by ScienCell (1 mentions)
Hepg2, by Thermo Fisher Scientific (1 mentions)
Hepg2, by American Type Culture Collection (1 mentions)
Serum free keratinocyte growth medium, by Thermo Fisher Scientific (1 mentions)
Recombinant human il 17, by Thermo Fisher Scientific (1 mentions)
Recombinant human ifn γ, by Thermo Fisher Scientific (1 mentions)
Anti cd3 antibody, by Thermo Fisher Scientific (1 mentions)
Str profiling, by Procell (1 mentions)
7 protocols using recombinant human il 22
1
IL-22 and PDGF-BB Signaling in Human Bronchial SMCs
2
Synovial Fluid Biomarkers in IJD
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Dkk1, sFRP3/FRZB, and IL-22 concentrations were measured directly in the synovial fluid (n=19; 13 RA, 3 PsA, and 3 AS) and in FLS culture supernatants, following the manufacturer's protocols (R&D systems, ref.: DY1906; DY192; DY210; DY317; and DY782). A pilot assay (n=3) for defining the optimal dose and time-to-collect was performed using one sample source from each IJD, none of them using synthetic or biological disease-modifying antirheumatic drugs, but only using non-steroidal anti-inflammatory drugs (NSAIDs) and corticosteroids (2.5-10 mg of prednisone/day). The FLS culture supernatant was collected at 24, 48, and 72 h after stimulus with recombinant human TNF-α (Peprotech, USA, ref.: 300-01A) or recombinant human IL-17A (Peprotech, ref.: 200-17), both at concentrations of 1, 10, 50, or 100 ng/mL, or recombinant human IL-22 (Peprotech, ref.: 200-22) at doses of 1, 10, 100, or 200 ng/mL. There were controls (both cells supernatants without stimulation and culture medium without cells) for all samples. Based on the results of this pilot study, the concentrations of sFRP3 were measured in a larger sample (n=11: 5 RA, 3 PsA, and 3 AS), 24 h after stimulus with IL-22, TNF-α, or IL-17, all at 10 ng/mL. All experiments were performed in three complete and independent repetitions.
3
Investigating HSV-2 Infection and Apoptosis
4
IL-22 Stimulation and STAT3 Immunoblotting
5
Culturing Hepatocyte Cell Lines for IL-22 Assays
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Human hepatoma cell line Huh7 was purchased from the Japanese Collection of Research Bioresources cell bank (JCRB0403; Osaka, Japan), and the human hepatoma cell line HepG2 was purchased from the American Type Culture Collection (HB‐8065; Manassas, VA). The Huh7 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) 1 g/L glucose (Gibco 31885‐023) supplemented with 10% fetal calf serum and 1% penicillin/streptomycin. HepG2 cells were cultured in DMEM 4.5 g/L glucose (Gibco 41965‐039). Recombinant human IL‐22 (Peprotech, Rocky Hill, NJ) and Recombinant human IL‐22BP/IL‐22RA2 protein (Sino Biological, Wayne, PA) were used for in vitro assays. Primary human hepatocytes were purchased from ScienCell Research Laboratories (Carlsbad, CA) and cultured in hepatocyte medium on poly‐l‐lysine‐coated plates according to the manufacturer’s instructions. Cell viability was assessed using the WST‐1 Cell Proliferation Reagent from Takara Bio (Kusatsu, Shiga, Japan).
6
Keratinocyte Culture and Cytokine Stimulation
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NHKs were extracted from prepuces obtained from healthy individuals who accepted circumcision. Informed consent was obtained from all donors. Keratinocytes were cultured in the serum-free keratinocyte growth medium (Gibco), and the second- or third-passage keratinocytes were used in all experiments. Cytokine cocktail used for stimulating NHKs was composed of recombinant human TNF-α (50 ng/ml), recombinant human IFN-γ (10 ng/ml), recombinant human IL-17 (25 ng/ml), and recombinant human IL-22 (25 ng/ml) (Peprotech, Rocky Hill City, CT, USA). Peripheral blood mononuclear cells (PBMCs) were obtained from psoriasis patients with informed consent and cultured in Modified Medium RPMI 1640 (HyClone, Logan, UT, USA) with 10% fetal bovine serum (Sijiqing, Hangzhou, China) before subsequent transwell assay. Each experiment was repeatedly performed using the cells at least from three different sources.
7
CD4+ T Cell Activation and NSCLC IL-22 Modulation
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CD4+ T cells were seeded into 24-well plates at a concentration of 106/ml, and were incubated in RPMI 1640 supplemented with 10% of heat-inactivated FBS at 37°C under 5% CO2 environment. Cells were stimulated by anti-CD3 antibody (eBioscience, Thermo Fisher, San Diego, CA, U.S.A.; final concentration, 1 μg/ml), with or without Notch signaling inhibitor, γ-secretase inhibitor (GSI) LY-411575 (Adooq, Irvine, CA, U.S.A.; final concentration, 1 μM) for 96 h. Lung adenocarcinoma cell line A549 was used for the study of direct IL-22 modulatory function to NSCLC. A549 cells were confirmed by STR profiling (Procell Life Science & Technology, Wuhan, Hubei Province, China; see Supplementary data). Confirmed A549 cells were cultured in DMEM containing 10% of FBS in the presence or absence of recombinant human IL-22 (Peprotech, Rocky Hill, NJ, U.S.A.; final concentration, 1 μg/ml) for 6 h. Cells and supernatants were harvested for further studies.
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