Serum from broilers was collected from a total of 36 chickens (three from each pen, nine for each treatment) following the methods described by Chuang et al. [11 (link)]. Briefly, chicken blood was collected and stored at 4 °C in a refrigerator R2551HS (TECO Electric and Machinery Co., Ltd., Taipei, Taiwan) for 4–5 h. Blood samples were centrifuged at 3000 rpm× g for 10 min in order to separate the blood cells and the serum, and the serum was stored at −20 °C in a refrigerator before being analyzed. The concentration of tumor necrosis factor-alpha (TNF-α), glutathione peroxidase (Gpx), and superoxidase dismutase (SOD) in broiler serum were measured (TNF-α, Gpx, and SOD were from Cayman Chemical Co., Ltd. Ann Arbor, MI, USA). All the methods for analyzing these compounds followed the manufacturer’s protocol. Other serum characteristics were measured with an automatic biochemical analyzer (Hitachi, 7150 auto-analyzer, Tokyo, Japan).
Tnf α
Manufactured by Cayman Chemical
Sourced in United States, China
TNF-α is a recombinant human tumor necrosis factor alpha (TNF-α) protein. TNF-α is a pro-inflammatory cytokine that plays a central role in the immune response and inflammation.
Automatically generated - may contain errors
Lab products found in correlation
Tnf α, by R&D Systems (4 mentions)
Il 1β, by Shanghai Enzyme-linked (3 mentions)
Il 10, by Shanghai Enzyme-linked (3 mentions)
Ifn γ, by GenStar (2 mentions)
Scd14, by R&D Systems (2 mentions)
I fabp, by R&D Systems (2 mentions)
Imark microplate reader, by Bio-Rad (2 mentions)
Tnf α, by Merck Group (2 mentions)
Mrp300, by R&D Systems (2 mentions)
Mob00b, by R&D Systems (2 mentions)
Rab0055, by Merck Group (2 mentions)
Microplate reader, by Thermo Fisher Scientific (2 mentions)
Trizol, by Thermo Fisher Scientific (2 mentions)
7150 autoanalyzer, by Hitachi (1 mentions)
Soluble p selectin, by R&D Systems (1 mentions)
Il 1β, by R&D Systems (1 mentions)
Elisa assay kit, by Cayman Chemical (1 mentions)
Interleukin 6 (il 6), by R&D Systems (1 mentions)
Mcp 1, by R&D Systems (1 mentions)
Il 10, by R&D Systems (1 mentions)
22 protocols using tnf α
1
Broiler Serum Biomarker Quantification
2
Modulation of TNF-α-Induced Responses by GPR40
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The NRK52E cells (American Type Culture Collection, Manassas, VA, USA) were treated with TNF-α (20 ng/mL; R&D Systems, Minneapolis, MN, USA) for 24 h in the presence or absence of the GPR40 agonist, GW9508 (10 μM; Cayman Chemical, Ann Arbor, MI, USA), for 1 h prior to the addition of TNF-α, and then harvested for further analysis. The control cells were treated with the vehicle (dimethyl sulfoxide). The NRK52E cells were also treated with TNF-α with or without a 1 h pretreatment with the GPR40 antagonist, GW1100 (10 μM; Cayman Chemical), or GPR40 small interfering (si)RNA (50 nM; ON-TARGET plus Rat Ffar1 (Gene ID: 266607), Item No. L-080051-02-0010, Dharmacon, Lafayette, CO, USA).
3
Venous Effluent Biomarker Dynamics
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The biochemical markers measured were plasma P-selectin, TNF-α, and MPO levels in the venous effluent in the control and insulin groups. Loco-regional sampling was performed at 0, +2, +4, +6, and +12 hours post reperfusion to establish the temporal evolution of P-selectin, TNF, and MPO in the venous effluent.
ELISA was used to measure plasma MPO (ng/mL; R&D Systems Inc., Minneapolis, MN, USA), soluble P-selectin (ng/mL; R&D Systems Inc.), and TNF-α (pg/mL; Cayman Chemical Company, Ann Arbor, MI, USA) levels.
ELISA was used to measure plasma MPO (ng/mL; R&D Systems Inc., Minneapolis, MN, USA), soluble P-selectin (ng/mL; R&D Systems Inc.), and TNF-α (pg/mL; Cayman Chemical Company, Ann Arbor, MI, USA) levels.
4
Profiling Immune and Gut Biomarkers in Clinical Cohorts
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Fasting peripheral blood samples were collected from all the subjects at 8:00 a.m. Blood cell count and liver function were examined routinely. The protein levels of indicators assessed by enzyme-linked immunosorbent assay (ELISA) in this study involved CRP (#E007462, 3ABio, Shanghai, China), IL-6 (#E000482, 3ABio, Shanghai, China), IL-1β (#E001772, 3ABio, Shanghai, China), IL-4 (#DG10308H, Dogesce, Beijing, China), IL-10 (#DG10495H, Dogesce, Beijing, China), IL-17 (#DG10431H, Dogesce, Beijing, China), IFN-γ (#C608-01, GenStar, Beijing, China), TNF-α (#489204, Cayman, Michigan, USA), TGF-β (#DG10113H, Dogesce, Beijing, China); leaky gut-related biomarkers[intestinal fatty acid-binding protein (I-FABP, #DFBP20, R&D Systems, Minnesota, USA) and Claudin-3 (#abx250611, Abbexa, Cambridge, UK)]; BT-related biomarkers[LPS (#DG11072H, Dogesce, Beijing, China), soluble CD14 (sCD14, #DC140, R&D Systems, Minnesota, USA), and endotoxin core antibody (EndoCAb, #E013362, 3ABio, Shanghai, China)]. Assays were performed according to the specifications of the manufacturer and the detection limits were in line with the instructions of the manufacturer. Each serum sample was measured in duplicate. All the plates were read by the I Mark™ Micro plate Reader (Bio-Rad, Hercules, California, United States).
5
Plasma Biomarkers in Insomnia Patients
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Plasma melatonin concentrations of both IDD patients and healthy controls were measured using human melatonin ELISA Kit (Cat#: 10,698, Glory Science, USA). IL-1β, IL6 and TNF-α levels in the plasma were measured using IL-1β (Cat#:QLB00B, R&D, USA), IL-6 (Cat#: RJ11850, Renjie Bio, China) and TNF-α (589,201–480, Cayman, USA) ELISA kit. All the measurements were conducted according to the manufacturer’s protocols. Three repeats for all the samples measurement were adopted and the average value was used in final analyses.
6
Equine Synovial Fluid ELISA
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Enzyme‐linked immunosorbent assays (ELISA) were run using commercially available kits validated for equine synovial fluid for IL‐1β (R&D Systems), TNF‐α (R&D Systems), and prostaglandin E2 (PGE2; Cayman Chemical) per manufacturer instructions. Samples were run in duplicate against a standard curve. Calculations of cytokine concentration from raw optical density data were performed using either proprietary (Cayman Chemical; PGE2) or freely available (elisaanalysis.com; IL‐1β, TNF‐α) software.
7
Cytokine Quantification by ELISA
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According to the manufacturer’s protocol, the levels of IL-1β (Shanghai Enzyme-Linked Biotechnology, cat#ml301814), TNF-α (Cayman Chemical, cat#500850), and IL-10 (Shanghai EnzymeLinked Biotechnology, cat#ml037873) were detected by indicated ELISA assay kits (Cayman Chemical, Michigan, USA). The absorbance value of each well was detected at 450 nm using the Microplate Reader (Thermo Scientific, NY, USA).
8
Resolvin Effects on Keratinocyte Inflammatory Response
9
Profiling Blood Biomarkers in EV-A71 Infection
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Peripheral blood samples were collected from all the subjects. Blood cell count and liver function were routinely examined. The protein levels of indicators assessed by enzyme-linked immunosorbent assay (ELISA) in present study involved CRP (#E007462, 3ABio), IL-1β (#E001772, 3ABio), IL-6 (#E000482, 3ABio), IFN-γ (#C608-01, GenStar), TNF-α (#489204, Cayman), LPS (#DG11072H, Dogesce), I-FABP (#DFBP20, R&D Systems), Claudin-3 (#abx250611, Abbexa), sCD14 (#DC140, R&D Systems) and EndoCAb (#E013362, 3ABio). Human EV-A71 VP1 protein ELISA kit (#MM-13481H2, MeiMian) was applied to detect the protein level of EV-A71 VP1 from blood serum. Assays were performed according to the manufacturer’s specifications and the detection limits were in line with the manufacturer’s instructions. All the plates were read by the I Mark™ Micro plate Reader (BIO-RAD).
10
Quantifying Inflammatory Mediators in Cell Culture
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Commercial kits were used to analyze prostaglandin 2 (PGE2) (500141) (Cayman Chemical), TNF-α (DY008), monocytes chemoattractant protein-1 (MCP-1) (DY479-05), interleukin-10 (IL-10) (DY417-05), IL-12 (DY419-05) and IL-6 (DY406-05) (R&D Systems), following the manufacturer’s instructions and according Grancieri, Martino and Gonzalez de Mejia [25 (link)]. The cell culture supernatants were diluted 1:5 (v/v, sample: buffer) for experiments I and II and 1:25 (v/v, sample: buffer) for experiments III and IV. The amount of PGE2 (y = −0.2766x + 0.3636, R2 = 0.99), TNF-α (y = 0.7991x − 2.0792, R2 = 0.99), MCP-1 (y = 0.7074x − 1.5536, R2 = 0.98), IL-6 (y = 0.7681x − 2.4798, R2 = 0.99); IL-10 (y = 0.7159x − 1.2982, R2 = 0.99), IL-12 (y = 0.7433x − 1.8984, R2 = 0.99) were calculated using log10, including their respective standard curves that were run at the same time as treatments. Absorbance was determined at 450 nm and results were expressed in pg/mL.
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