The HTIW under investigation are all in commercial production, but the manipulations were done specifically for this study by Morgan Advanced Materials (Bromborough, UK) and were provided to Heriot-Watt University in a coded form to allow in vitro experiments to be blinded. DQ12 quartz was donated by the Institute of Occupational Medicine (IOM) (Edinburgh, UK) (DQ12 was included in this study as a positive control for effects pertaining to crystalline silica); TiO2 was from Tioxide, UK; nanoparticle carbon black (NPCB) was obtained from Degussa (Printex 90). rhTNF-α was from Immunotools (Friesoythe, Germany); AlamarBlue reagent, Gibco™ Penicillin-Streptomycin, RPMI 1640, phenol red-free MEM were from Thermo Fisher (Paisley, UK); QCL-1000 Endpoint Chromogenic LAL Assay was from Lonza (Slough, UK); Proteome Profiler Cytokine Array Kit, Magnetic Luminex Screening Assay, and DuoSet ELISA kits were from R&D Systems (Abingdon, UK); 1-hydroxyl-2,2,6,6-tetramethyl-4-oxo-piperidine (Tempone-H) was from Enzo Life Sciences (Exeter, UK). All other substances used were obtained from Sigma-Aldrich (Poole, UK).
Rhtnf α
Manufactured by Immunotools
Sourced in Germany, United Kingdom
RhTNF-α is a recombinant human tumor necrosis factor alpha (TNF-α) protein. TNF-α is a cytokine that plays a central role in inflammation and immune system regulation.
Automatically generated - may contain errors
Lab products found in correlation
Gibco penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Duoset elisa kit, by R&D Systems (1 mentions)
Phenol red free mem, by Thermo Fisher Scientific (1 mentions)
Printex 90, by Evonik (1 mentions)
Qcl 1000 endpoint chromogenic lal assay, by Lonza (1 mentions)
Alamarblue reagent, by Thermo Fisher Scientific (1 mentions)
Proteome profiler cytokine array kit, by R&D Systems (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Magnetic luminex screening assay, by R&D Systems (1 mentions)
Celltiter blue ctb cell viability assay, by Promega (1 mentions)
Camptothecin cpt, by Abcam (1 mentions)
Il 8 elisa kit, by Thermo Fisher Scientific (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Revertaid h minus m mulv reverse transcriptase, by Thermo Fisher Scientific (1 mentions)
Sybr green supermix, by Bio-Rad (1 mentions)
Rebif, by Merck Group (1 mentions)
Lymphocyte separating medium, by Corning (1 mentions)
Aim 5 medium, by Thermo Fisher Scientific (1 mentions)
Human ab serum, by Merck Group (1 mentions)
Rhifn γ, by Immunotools (1 mentions)
4 protocols using rhtnf α
1
Cytotoxicity and Inflammatory Profiling of HTIW
2
Characterization of Copper Oxide Nanoparticles
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CuO NPs were obtained from Intrinsiq Materials Ltd (Farnborough, UK) and were supplied by the Nanovalid consortium (http://www.nanovalid.eu/ ). Acetonitrile (ACN) for LC-MS was purchased from VWR (Radnor, PA, USA). High-purity water (H2O) was produced using a Milli-Q Integral three purification system from Merck Millipore (Darmstadt, Germany). Standard substances used for identification were obtained from Merck (amino acids). Staurosporine (STS) was purchased from Proteinkinase.de (Kassel, Germany), camptothecin (CPT) from Abcam (Cambridge, UK), and rhTNF-α from Immunotools (Friesoythe, Germany). SYBR Green Supermix was purchased from Bio-Rad (Munich, Germany), RevertAid HMinus M-MulV reverse transcriptase from Fermentas (St. Leon-Roth, Germany), TRIzol reagent from Invitrogen, IL-8 ELISA kits from PeproTech, while Celltiter-Blue® (CTB) Cell Viability Assay was purchased from Promega (Madison, WI, USA), foetal calf serum (FCS) from PAA (Pasching, Austria). All other substances used were obtained from Sigma-Aldrich (St. Louis, MO, USA).
3
Cytokine-Induced Chemokine Profiling
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4
Generation of Activated Dendritic Cells
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The peripheral blood mononuclear cells (PBMCs) were isolated from 30 ml of HLA-A2+ healthy donor blood with written consent under Siriraj Institutional Review Board ethical approval (COA no. Si 580/2018) by density centrifugation at 800 × g for 20 min at RT in lymphocyte separating medium (Corning Life Sciences). The monocytes were isolated and incubated for 1 h at 37°C. The non-adherence cells were collected and cryopreserved in human AB serum (MilliporeSigma) containing 10% dimethyl sulfoxide until use. The monocytes were transduced with IRFP-SmartDC or MSLN-SmartDC at 75 multiplicity of infection (MOI) together with 10 µg/ml protamine sulfate in AIM-V medium (Invitrogen; Thermo Fisher Scientific, Inc.). On day 5 post-transduction, 1 µg/ml of recombinant human RPS3 (cat. no. NBP2-90977; Novus Biologicals, LLC) was added. Monocytes were cultured in 100 ng/ml of rhGM-CSF (cat. no. 11343125; ImmunoTools GmbH) and 50 ng/ml of rhIL-4 (cat. no. 11340045; ImmunoTools GmbH) for five days and treated with 100 ng/ml of rhIFN-γ (cat. no. 11343536; ImmunoTools GmbH) and rhTNF-α (cat. no. 11343015; ImmunoTools GmbH) for additional 48 h served as positive control or conventional DC (conv. DC). All DCs were harvested on day 7 to check the activated characters.
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