Mouse il 6 quantikine elisa kit

Manufactured by R&D Systems

Sourced in United States, Germany, Canada, United Kingdom

The Mouse IL-6 Quantikine ELISA Kit is a quantitative sandwich enzyme immunoassay designed to measure mouse interleukin 6 (IL-6) levels in cell culture supernates, serum, and plasma. The kit utilizes a microplate pre-coated with a monoclonal antibody specific for mouse IL-6.

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101 protocols using mouse il 6 quantikine elisa kit

Muscle Hypertrophy Biomarkers Quantification

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We measured levels of interleukin (IL)-6, IL-15, IL-8 and IL-1β using a Mouse IL-6
Quantikine ELISA Kit (R&D Systems, Inc. USA), Mouse IL-15 ELISA Kit (Abcam plc, Cambridge,
UK), Mouse CXCL2/MIP-2 Quantikine ELISA Kit (R&D Systems, Inc. USA) and Mouse IL-1 beta
ELISA Kit (Abcam plc, Cambridge, UK). For analyses of muscular hypertrophy, we undertook
western blotting using antibody against the myogenic determination (MyoD) gene (H2107; Santa
Cruz Biotechnology, Santa Cruz, CA, USA) and measured the detected bands with ImageJ (National
Institutes of Health, Bethesda, MD, USA).

Kito T., Nishii K., Yao R., Teranishi T., Sugiyama T., Sakai K., Matsubara M, & Yamada K. (2019). Effect of shaking and vibration stimulation on lumbar vertebrae in ovariectomized mice. Fujita Medical Journal, 5(3), 57-62.

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Inflammatory Pathway Activation Assays

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InVivoMab anti-mouse Ly6G (CAT#: BE0075-1) was purchased from Bio X Cell (Lebanon, NH, USA). Compound C (CpC, CAT#: 171260), LPS (E. coli O111:B4, CAT#: L4391), STO-609 (STO, CAT#: 570250) and lucigenin (CAT#: M8010) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Mouse interleukin-1β (IL-1β)/IL-1F2 Quantikine ELISA Kit (CAT#: MLB00C) and Mouse IL-6 Quantikine ELISA Kit (CAT#: M6000B) were purchased from R&D Systems, Inc. (Minneapolis, MN, Canada). MojoSort™ Human Pan Monocyte Isolation Kit (CAT#: 480060) was purchased from BioLegend (USA). Human Macrophage Colony-Stimulating Factor (M-CSF) Recombinant Protein (CAT#: RP-8643), Pierce™ BCA Protein Assay Kit (CAT#: 23227) and Amplex Red Hydrogen Peroxide⁄Peroxidase Assay Kit (CAT#: A22188) were purchased from Invitrogen (Carlsbad, CA, USA). Lactate Dehydrogenase (LDH) Assay Kit (CAT#: A020-2), Myeloperoxidase (MPO) Assay Kit (CAT#: A044-1-1), ROS Assay Kit (CAT#: E004-1-1), Malondialdehyde (MDA) Assay Kit (CAT#: A003-1-1), 4-Hydroxynonenal ELISA Kit (CAT#: H268), Protein Carbonyl Assay Kit (CAT#: A087-1-1), Total Anti-oxidant Capacity Assay Kit (CAT#: A015-2-1) and Superoxide Dismutase (SOD) Assay Kit (CAT#: A001-3-2) were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). TransAM® NF-κB p65 Kit was purchased from Active Motif (Carlsbad, CA, USA).

Chen R., Cao C., Liu H., Jiang W., Pan R., He H., Ding K, & Meng Q. (2022). Macrophage Sprouty4 deficiency diminishes sepsis-induced acute lung injury in mice. Redox Biology, 58, 102513.

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Quantifying IL-6 Secretion in Cells and Serum

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For investigation of IL-6 secretion by HuH-7 cell or in mouse serum, we used the Human IL-6 Quantikine ELISA Kit and the Mouse IL-6 Quantikine ELISA Kit (both R&D Systems), respectively. Procedures were performed according to the manufacturer's protocols. Triplicates of each sample were measured. For cell culture supernatants, two biological replicates were performed.

Feng T., Dzieran J., Yuan X., Dropmann A., Maass T., Teufel A., Marhenke S., Gaiser T., Rückert F., Kleiter I., Kanzler S., Ebert M.P., Vogel A., ten Dijke P., Dooley S, & Meindl-Beinker N.M. (2017). Hepatocyte-specific Smad7 deletion accelerates DEN-induced HCC via activation of STAT3 signaling in mice. Oncogenesis, 6(1), e294-.

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Quantifying Cytokine Responses in Viral Infections

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Total RNA was purified using RNeasy kit (QIAGEN) and DNase-treated RNA was reverse transcribed with PrimeScript RT reagent kit (Takara-Bio) for quantitative real-time PCR using Thermal Cycler Dice Real Time System (Takara-Bio). The relative abundance of mRNA was obtained by the ΔΔC_T method, using GAPDH for normalization. Culture media from infected cells or control cells were analyzed for cytokine production using mouse IL-6 Quanti-kine ELISA kit (R&D systems) and mouse IFNμ ELISA kit (PBL Assay Science).

Komuro A., Homma Y., Negoro T., Barber G.N, & Horvath C.M. (2016). The TAR-RNA binding protein is required for immunoresponses triggered by Cardiovirus infection. Biochemical and biophysical research communications, 480(2), 187-193.

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Quantifying Inflammatory Mediators in Colonic Tissue

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Levels of MPO and IL‐6 in colonic tissue were measured using a mouse MPO ELISA kit (HK210; Hycult Biotechnology, Uden, The Netherlands), and a mouse IL‐6 Quantikine ELISA kit (R&D Systems Inc., Minneapolis, MN, USA), respectively, according to manufacturer’s instructions. To analyse TNFα, total RNA was extracted from colonic tissues using TRIzol^TM Reagent (Invitrogen, Carlsbad, CA, USA), according to manufacturer’s instructions, and quantified using a NanoDrop ND‐1000 (Thermo Fisher Scientific, Waltham, MA, USA). Complementary DNA (cDNA) was synthesized using a High‐Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Carlsbad, CA, USA) and was analysed using quantitative real‐time (qRT) PCR, with TNFα specific primers (Table S2) and Power SYBR^TM Green PCR Master Mix (Thermo Fisher Scientific), in a ViiA 7 instrument (Thermo Fisher Scientific). Every assay was performed in triplicates.

Park Y., Kim T., Ham J., Choi J., Lee H., Yeon Y.J., Choi S.I., Kim N., Kim Y, & Seok Y. (2021). Physiological activity of E. coli engineered to produce butyric acid. Microbial Biotechnology, 15(3), 832-843.

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Terrein Modulates Cytokine Secretion

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The RAW 264.7 cells were plated at 5 × 10⁵ cells/mL and stimulated with LPS (200 ng/mL) in the presence or absence of terrein (1, 3, 10, or 30 μM) for 24 h. The culture supernatants were collected and the amount of cytokine was determined using a mouse IL-1β Quantikine ELISA kit or mouse IL-6 Quantikine ELISA kit (R&D Systems, Inc., Minneapolis, MN, USA) according to the manufacturer’s instructions.

Yoon Y.D., Lee M.Y., Choi B.J., Lee C.W., Lee H., Kwon J.H., Yang J.W, & Kang J.S. (2022). Protection against Lipopolysaccharide-Induced Endotoxemia by Terrein Is Mediated by Blocking Interleukin-1β and Interleukin-6 Production. Pharmaceuticals, 15(11), 1429.

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Cytokine Profiling in Murine Lung and Serum

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Blood samples were obtained by cardiac puncture and collected in heparinized tubes [36 (link),60 (link)]. BAL samples were obtained according to the technique developed in the Laboratory Immunobiotechnology of CERELA-CONICET (San Miguel de Tucuman, Argentina) [38 (link),61 (link)]. The trachea was exposed and intubated with a catheter, and then two sequential bronchoalveolar lavages were performed on each mouse by injecting sterile PBS. The recovered fluid was centrifuged for 10 min at 900× g and the fluid frozen at −70 °C for subsequent cytokine determinations. Tumor necrosis factor alpha (TNF-α), interferon gamma (IFN-γ), interleukin 10 (IL-10), IL-1β, IL-6, IL-17, KC, and monocyte chemoattractant protein 1 (MCP-1) concentrations were determined in serum and BAL samples using commercial ELISA kits. IFN-γ (Mouse IFN-gamma Quantikine ELISA Kit, sensitivity: 2 pg/mL), IL-10 (Mouse IL-10 Quantikine ELISA Kit, sensitivity: 5.2 pg/mL), IL-1β (Mouse IL-1β DuoSet ELISA, sensitivity: 1.5 pg/mL), IL-6 (Mouse IL-6 Quantikine ELISA Kit, sensitivity: 1.8 pg/mL), and IL-17 (Mouse IL-17 Quantikine ELISA Kit, sensitivity: 4.7 pg/mL) from R&D Systems (USA). TNF-α (Mouse TNF alpha ELISA Kit, sensitivity: 9.1 pg/mL), MCP-1 (Mouse MCP1 ELISA Kit, sensitivity: 0.487 pg/mL), and KC (Mouse KC ELISA Kit, sensitivity: 1.95 pg/mL) kits were obtained from Abcam (Cambridge, UK).

Albarracin L., Ortiz Moyano R., Vargas J.M., Andrade B.G., Cortez Zamar J., Dentice Maidana S., Fukuyama K., Kurata S., Jure M.Á., Kitazawa H, & Villena J. (2022). Genomic and Immunological Characterization of Hypermucoviscous Carbapenem-Resistant Klebsiella pneumoniae ST25 Isolates from Northwest Argentina. International Journal of Molecular Sciences, 23(13), 7361.

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Mouse Cytokine ELISA Assays

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LBIS^® Anti-dsDNA-Mouse ELISA Kit (631-02699, Amresco, Radnor, PA, USA), Mouse C3 ELISA Kit (ab263884, Abcam), Mouse IL-1 beta/IL-1F2 Quantikine ELISA Kit (MLB00C, R&D Systems, Minneapolis, MN, USA), Mouse IL-6 Quantikine ELISA Kit (M6000B, R&D Systems), and TNF-α (Mouse) ELISA Kit (K1051, BioVision, Inc., Exton, PA, USA) were used.

Qi H., Zheng Z, & Liu Q. (2023). Activation of BZW1 by CEBPB in macrophages promotes eIF2α phosphorylation-mediated metabolic reprogramming and endoplasmic reticulum stress in MRL/lpr lupus-prone mice. Cellular & Molecular Biology Letters, 28, 79.

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Quantification of Inflammatory Cytokines

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The levels of IL11, IL6, CCL2, and CCL5 in equal volumes of cell culture media were quantified using Human IL11 Quantikine ELISA kit (D1100; R&D Systems), Human IL-6 Quantikine ELISA Kit (D6050; R&D Systems), Human CCL2/MCP-1 Quantikine ELISA Kit (DCP00; R&D Systems), and Human CCL5/RANTES Quantikine ELISA Kit (DRN00B; R&D Systems), respectively. The levels of IL11, IL6, gp130, and IL11RA in mouse serum were quantified using Mouse IL11 DuoSet ELISA (DY418; R&D Systems), Mouse IL6 Quantikine ELISA Kit (M6000B; R&D Systems), Mouse Interleukin 11 Receptor Alpha (IL11Ra) ELISA Kit (MBS452535; MyBioSource), and Mouse gp130 DuoSet ELISA (DY468, R&D Systems), respectively. All ELISA assays were performed according to the manufacturer’s protocol.

Dong J., Viswanathan S., Adami E., Singh B.K., Chothani S.P., Ng B., Lim W.W., Zhou J., Tripathi M., Ko N.S., Shekeran S.G., Tan J., Lim S.Y., Wang M., Lio P.M., Yen P.M., Schafer S., Cook S.A, & Widjaja A.A. (2021). Hepatocyte-specific IL11 cis-signaling drives lipotoxicity and underlies the transition from NAFLD to NASH. Nature Communications, 12, 66.

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Cytokine Quantification in Cell Culture

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Cell culture supernatants and SNc were assayed for IL-1β (Mouse IL-1 beta/IL-1F2 Quantikine ELISA Kit, MLB00C), TNF-α (Mouse TNF-alpha Quantikine ELISA Kit, MTA00B), IL-6 (Mouse IL-6 Quantikine ELISA Kit, M6000B), and IL-12 (Mouse IL-12 p70 Quantikine ELISA Kit, M1270) with ELISA kits from R&D Systems according to the manufacturer’s instructions.

Du R.H., Zhou Y., Xia M.L., Lu M., Ding J.H, & Hu G. (2018). α-Synuclein disrupts the anti-inflammatory role of Drd2 via interfering β-arrestin2-TAB1 interaction in astrocytes. Journal of Neuroinflammation, 15, 258.

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