Colon tissue was homogenized in 1 mL PBS and centrifuged (13,000 rpm for 20 min at 4 °C). Supernatant was filtered (0.22 μm), and protein concentration was determined by Bradford assay. The presence of cytokines was analyzed in supernatant fractions with mouse antibody arrays Q1 (ABIN625794; RayBiotech) following the manufacturer’s instructions. ELISA kits were used to quantify IL-22, IL-13, IL-10, TGF-β, IL-23, IL-21, IL-6, and TNF-α (RayBiotech).
Il 13
Manufactured by RayBiotech
Sourced in United States
IL-13 is a cytokine that plays a role in immune and inflammatory responses. It is involved in the regulation of gene expression and cellular functions.
Automatically generated - may contain errors
Lab products found in correlation
Interleukin 6 (il 6), by RayBiotech (4 mentions)
Interleukin 4 (il 4), by RayBiotech (4 mentions)
Tnf α, by RayBiotech (3 mentions)
Il 10, by RayBiotech (3 mentions)
Mcp 1, by RayBiotech (2 mentions)
Il 1β, by RayBiotech (2 mentions)
Interleukin 2 (il 2), by RayBiotech (2 mentions)
Il 21, by RayBiotech (1 mentions)
Il 18, by RayBiotech (1 mentions)
Mini protean 3 cell, by Bio-Rad (1 mentions)
Envision reagent, by Agilent Technologies (1 mentions)
20 r rg3, by Yuanye Bio-Technology (1 mentions)
Cell cycle kit, by BD (1 mentions)
Cyclophosphamide cp, by Merck Group (1 mentions)
Annexin 5 fitc, by BD (1 mentions)
Automated cell counter, by Beckman Coulter (1 mentions)
Vegf a, by RayBiotech (1 mentions)
Tgf β, by MyBioSource (1 mentions)
M csf, by RayBiotech (1 mentions)
M csf, by Apexbio (1 mentions)
6 protocols using il 13
1
Cytokine Profiling of Colon Tissue
2
Murine Asthma Model Protocol
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Ovalbumin (OVA) was purchased from Shanghai Junhao Chemical Co., Ltd. (Shanghai, China). Murine antibodies of NGF (diluted at 1:50) and TrkA (diluted at 1:100) were purchased from Abcam (Cambridge, UK). Enzyme-linked immunosorbent assay (ELISA) kits including TrkA, NGF, immunoglobulin (Ig) E, IL-13 and IL-18 were obtained from RayBiotech Inc. (Norcross, GA, USA). EnVision reagent was obtained from Dako; Agilent Technologies, Inc. (Santa Clara, CA, USA) and horseradish peroxidase (HRP) conjugated anti-mouse antibodies were obtained from Beyotime Institute of Biotechnology (Shanghai, China). Optical densities of samples were determined using an MK3 microplate reader (Labsystems Diagnostics, Vantaa, Finland). A Mini-PROTEAN 3 Cell (Bio-Rad Laboratories, Inc., Hercules, CA, USA) was used for electrophoresis. Dexamethasone (Dxm) acetate tablets (0.75 mg/tablet) were provided by the Shanghai Xinyi Pharm. Co., (Shanghai, China).
3
Quantification of Ginsenosides and Monosaccharides
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The reference substances of ginsenosides (nR1, Rg1, Re, pF11, Rf, Ra2, Rb1, Rc, Ro, Rb2, Rb3, Rd and 20(R)-Rg3) and monosaccharides (Glc, Ara, Gal, Man, GalA, Rha, Rib, ClcUA, Xyl and Fuc) were purchased from Shanghai Yuanye Biotechnology (Shanghai, China). The purity of these references was higher than 99.0% indicated by HPLC analysis. Cyclophosphamide (CP) was supplied by Sigma-Aldrich Company Ltd., (United States). Thrombopoietin (TPO), erythropoietin (EPO), and granulocyte-macrophage colony stimulating factor (GM-CSF) were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Quantibody®Array Glass Chip of IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-10, IL-13, IFNγ, TNF-α and MCP-1 were purchased from RayBiotech Life, Inc. (United States). FITC-Annexin-V and Cell Cycle kits were purchased from BD Biosciences Pharmingen (United States). The blood cell analysis reagent kit was purchased from IDEXX Laboratories Inc (United States). Protopanaxadiol saponins (PDS) and protopanaxatriol saponins (PTS) were supplied by Professor Chen Yanping in the College of Chemistry, Jilin University; PDS and PTS were LC-MS grade. The voucher specimen of PDS (S20190011) and PTS (S20190012) were deposited in the authors’ lab in Changchun University of Chinese Medicine (Changchun, China).
4
Plasma Biomarker Profiling Protocol
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Plasma samples were obtained by venipuncture after an overnight fasting. Glucose was analyzed by an automated analyzer (Hitachi Modular P800, Roche, Basel, Switzerland). Serum concentrations of triglycerides and free fatty acids (FFA) were determined by using commercially available kits (Infinity, Thermo Electron Corporation, Melbourne, Australia). The carcinoembryonic antigen (CEA), fibrinogen and high sensitivity C-reactive protein (CRP) concentrations were measured as previously reported [11 (link)]. White blood cell (WBC) count was determined using an automated cell counter (Beckman Coulter, Inc., Fullerton, CA, USA). Circulating levels of MAGP-1, IL-4, IL-6, IL-13, VEGFA, OPN (RayBiotech, Inc., Norcross, GA, USA) and TGF-β (Mybiosource, San Diego, CA, USA) were assessed by commercially available ELISA kits according to the manufacturer’s instructions. The intra- and inter-assay coefficients of variation were <10 and <12% for all analysed molecules.
5
Multiplex Analysis of Rat Synovial Inflammation
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Rats were euthanized via CO2 inhalation at three days or twenty-one days post-surgery for both the naïve group and the MMT surgery group. Synovial fluid was collected by first injecting 100 μl of saline intra-articularly using a 30 gauge insulin syringe, followed by aspirating approximately 50 to 100 μl of the synovial fluid and saline using the same syringe. Synovial fluid was analyzed using the Quantibody® Rat Inflammation Array 1, a multiplex ELISA kit that quantitatively measured 10 rat inflammatory factors: IFNγ, IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-10, IL-13, monocyte chemoattractant protein-1 (MCP-1), and TNFα (RayBiotech, Norcross, GA, USA).
6
Isolation and Polarization of Rat Bone Marrow-Derived Macrophages
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The femur was isolated from euthanized Wistar male rats and marrow was flushed into a cold PBS solution containing 2% fetal bovine serum (FBS). Mechanically dissociated cells were incubated on ice with three volume of 0.8% NH4Cl solution to remove red blood cells. 106 cells/cm2 were seeded in tissue culture plates (Corning Constar) and cultured in Iscove's Modified Dulbecco's Medium (IMDM, Gibco) supplemented with 10% FBS, 1% v/v penicillin-streptomycin (Life Technologies) 10ng/ml M-CSF (RayBiotech). After 7 days, bone marrow-derived macrophages (MΦ) were polarized to inflammatory M1 with IMDM, 1% v/v penicillin-streptomycin, 10ng/ml M-CSF supplemented with 100ng/ml TNFα (ApexBio) and 100ng/ml IFNγ (RayBiotech). The non-inflammatory BMDM-M2 profile was induced by supplementing the culture medium with 20ng/ml IL-4, 20ng/ml IL-10 and 20ng/ml IL13 (RayBiotech).
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