To confirm the delayed cytokine expression with ProLNG-001, the level of tumor necrosis factor (TNF)-α in mouse plasma was measured. C57BL/6 mice were subcutaneously administered TBS, R848 (25 μg), or ProLNG-001 (140 μg). Mouse plasma was collected at various time points (0, 1, 3, 6, and 24 h) following administration. The plasma was separated from heparinized blood by centrifugation at 13,000× g rpm for 30 min, and the level of TNF-α in the isolated mouse plasma was analyzed using the BD OptEIA Mouse TNF (Mono/Mono) ELISA Set (BD Biosciences) following the manufacturer’s instructions.
Opteia mouse tnf mono mono elisa set
Manufactured by BD
Sourced in United States
The BD OptEIA Mouse TNF (Mono/Mono) ELISA Set is a laboratory product designed for the quantitative measurement of mouse tumor necrosis factor (TNF) in biological samples. It utilizes a sandwich enzyme-linked immunosorbent assay (ELISA) method to detect and quantify the target analyte.
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5 protocols using opteia mouse tnf mono mono elisa set
1
Delayed Cytokine Expression with ProLNG-001
2
Cytokine Profiling of Bone Marrow-Derived Macrophages
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BMDMs were plated at 50,000 cells/well, and cytokine concentrations in
cell supernatants were measured using the BD OptEIA Mouse IL-6 ELISA Set (BD
555240), BD OptEIA Mouse IL-12 (p40) ELISA Set (BD 555165), or BD OptEIA Mouse
TNF (Mono/Mono) ELISA Set (BD 555268) according to manufacturer
instructions.
cell supernatants were measured using the BD OptEIA Mouse IL-6 ELISA Set (BD
555240), BD OptEIA Mouse IL-12 (p40) ELISA Set (BD 555165), or BD OptEIA Mouse
TNF (Mono/Mono) ELISA Set (BD 555268) according to manufacturer
instructions.
3
Cytokine Profiling of Bone Marrow-Derived Macrophages
4
Quantification of Inflammatory Cytokines in Arthritis Models
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After all functional testing had been completed on day 20 in osteoarthritis or on day 19 in rheumatoid arthritis model, all animals were deeply anaesthetized with ketamine (100 mg/kg, i.p., Richter Gedeon Plc., Hungary) and xylazine (10 mg/kg, i.m., Lavet Ltd., Hungary); then the thoracic cavity was opened and 0.5–1 ml blood was collected from the left ventricle with a heparinized syringe into ice-cold polypropylene tubes containing EDTA (40 μl) and trasylol (20 μl). The collected blood was immediately centrifuged (1,000 g, 20 min, 4°C), and the resulting plasma was stored at -80°C. The concentrations of the inflammatory cytokines IL-1β and TNF-α were measured by ELISA using BD OptEIA Mouse IL-1β ELISA set and BD OptEIA Mouse TNF (Mono/Mono) ELISA Set (BD Biosciences, USA). Detection was performed by using Labsystem Multiscan RC plate reader (LabX Ltd., Canada).
5
Cytotoxicity Evaluation of Biomaterials
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The eventual cytotoxicity of the biomaterial was evaluated by measuring nitric oxide (NO), interleukin-1β (IL1β), and tumor necrosis factor α (TNFα) production using a model of macrophages in culture. Briefly, RAW264.7 macrophages were grown on the scaffolds or the tissue culture plates (control condition) in DMEM without phenol red-5% FBS. After 24, 48, and 72 h the supernatants were collected and evaluated for NO production by the Griess' assay. IL1β and TNFα production were evaluated by ELISA kits (BD OptEIA mouse IL-1β ELISA and BD OptEIA mouse TNF (Mono/Mono) ELISA set) in the conditioned media 2, 7, and 13 d of incubation.
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