Enzyme-linked immunosorbent assay (ELISA) kits for TNF-α, IL-1β and CCL2/MCP-1, were purchased from R&D Systems (Minneapolis, MN). ELISA kit for Dopamine was purchased from GenWay Bioteck Inc, San Diego, CA. Polyclonal anti-NF-κB p65 and cyclo-oxygenase-2 (COX-2) antibodies were obtained from Cell Signaling Technology (Danvers, MA) and Santa Cruz Biotechnology (Santa Cruz, CA). Secondary antibodies conjugated to horseradish peroxidase (Bio-Rad Laboratories, Hercules, CA), Rotamex Rotarod (Columbus Instruments, Columbus, OH). Male C57BL/6 mice (Harlan Sprague Dawley, Inc., Indianapolis, IN) and MPTP (Sigma, St. Louis, MO) were purchased.
Elisa kits for tnf α
Manufactured by R&D Systems
Sourced in United States, United Kingdom
ELISA kits for TNF-α are laboratory assays used to quantify the levels of tumor necrosis factor alpha (TNF-α) in biological samples. TNF-α is a cytokine that plays a role in inflammation and immune response. These kits provide a standardized and reliable method for measuring TNF-α concentration in research applications.
Automatically generated - may contain errors
Lab products found in correlation
Il 1β, by R&D Systems (4 mentions)
Lipopolysaccharides (lps), by Merck Group (3 mentions)
P jnk, by Cell Signaling Technology (2 mentions)
Interleukin 6 (il 6), by R&D Systems (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Mcp 1 ccl2, by R&D Systems (1 mentions)
Secondary antibodies conjugated to horseradish peroxidase, by Bio-Rad (1 mentions)
Rotamex rotarod, by Columbus Instruments (1 mentions)
C57bl 6 mice, by Merck Group (1 mentions)
Tnf α, by Enzo Life Sciences (1 mentions)
Complete lysis m buffer, by Roche (1 mentions)
Mikro dismembrator, by Sartorius (1 mentions)
Baclofen, by Bio-Techne (1 mentions)
Saclofen, by Bio-Techne (1 mentions)
Rantes ccl5, by R&D Systems (1 mentions)
Hmgb1 elisa kit, by Tecan (1 mentions)
Grp enzyme immunoassay eia, by Phoenix Pharmaceuticals (1 mentions)
Gaba elisa kit, by Aviva Systems Biology (1 mentions)
8 hydroxy 2 deoxyguanosine 8 ohdg elisa kit, by Abcam (1 mentions)
Elisa kit for ifn β, by PBL Assay Science (1 mentions)
26 protocols using elisa kits for tnf α
1
Comprehensive Neuroinflammation Measurement
2
Tissue Cytokine Quantification by ELISA
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To assess the cytokine concentration in the tissue, half of each tissue sample was pulverized with a mikro‐dismembrator (Sartorius, Goettingen, Germany), as described by van Dijk et al.26 To each sample 1 ml of EDTA‐free cOmplete Lysis‐M buffer (Roche, Almere, the Netherlands) was added, and samples were incubated overnight at 4°C and stored at −80°C. The cytokine concentrations in the tissue were measured with bovine specific ELISA kits for TNF‐α (detection limit: 125.0 pg/ml, R&D Systems, Abingdon, UK), IL‐1β (detection limit: 31.3 pg/ml, Thermo Fisher Scientific and MyBioSource, Uithoorn, the Netherlands), IL‐6 (detection limit: 78.1 pg/ml, Thermo Fisher Scientific), and IL‐8 detection limit: 8 pg/ml, Mabtech, Nacka Strand, Sweden).
Furthermore, the concentrations of TNF‐α, IL‐1β, IL‐6, IL‐8, and PGE2 (detection limit: 13.4 pg/ml, Enzo Life Sciences, Raamdonksveer, the Netherlands) were measured in the medium by ELISA. The concentrations were normalized to the initial sample wet weights to obtain the overall tissue response. If possible, standard curves were prepared in culture medium to account for medium effects. If this was not possible, the cytokine concentration in the medium was determined as control on the plate.
Furthermore, the concentrations of TNF‐α, IL‐1β, IL‐6, IL‐8, and PGE2 (detection limit: 13.4 pg/ml, Enzo Life Sciences, Raamdonksveer, the Netherlands) were measured in the medium by ELISA. The concentrations were normalized to the initial sample wet weights to obtain the overall tissue response. If possible, standard curves were prepared in culture medium to account for medium effects. If this was not possible, the cytokine concentration in the medium was determined as control on the plate.
3
Immunoassay Panel for Inflammatory Markers
4
Cellular Response to Inflammatory Stimuli
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Dulbecco’s modified Eagle medium (DMEM), fetal bovine serum (FBS), and penicillin-streptomycin (PS) were purchased from Invitrogen (Carlsbad, CA, USA). LPS, N-Monomethyl-l -arginine (NMMA), and AITC were purchased from Sigma-Aldrich (St. Louis, MO, USA). ELISA kits for TNF-α, IL-6 and NGF were bought from R&D Systems (Minneapolis, MN, USA), PGE-2 kit was purchased from cayman chemical (Ann Arbor, MI, USA). Primary and secondary antibodies against iNOS, COX2, ERK, pERK, JNK, pJNK, p38, pP38, Bax, Bcl-2, cleaved caspase-3, and tubulin were purchased from Cell Signaling (Beverly, MA, USA). All other chemicals and reagents were purchased from Sigma Chemical Co. (St. Louis, MO, USA).
5
Cytokine Quantification in Gastric Mucosa
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The gastric mucosal tissue homogenate obtained from the mucus model adhered to the gastric mucosa was centrifuged (10,000 rpm, 4 °C, 5 min) and the cytokines were detected in the supernatant using ELISA kits for TNF-α, IL-6, IL-1β, and IL-10 according to the manufacturer’s instructions at 450nm (R&D Systems, Minneapolis, MN, USA). The plates were read using a microplate reader SpectraMax i3 (Molecular Devices, San José, CA, USA).
6
Quantifying Immune Cytokine and Metabolic Profiles
7
Macrophage Cytokine Release Assay
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Mouse peritoneal activated macrophages were gained as described in the section cell culture. About, 4 × 105 cells per well were cultured in 96-well plates (TPP, Trasadingen, Switzerland). Cytokine release was assayed by an ELISA as described previously 27 (link). Briefly, cells were pre-treated with HCE for 30 min., and then cytokine production was stimulated by the addition of LPS from Escherichia coli (1 μg/ml) for 24 hrs. After cell lysis, the amount of synthesised pro-inflammatory cytokines was quantified in supernatants with commercially available ELISA kits for TNFα (R&D Systems), interleukin (IL)-6 (R&D Systems) and IL-β (R&D Systems) in accordance to the manufacturer's protocol.
8
Quantification of Cytokines and NAD Levels
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Cytokine concentrations in cell-free supernatants or total protein lysates (0.1% Triton X-100) were measured using ELISA kits for TNF-α (R&D Systems), IL-1β, and IL-6 (eBioscience). Standard curves were generated with a Four Parametric Logistic curve fit and data were analyzed using MyAssays Analysis Software Solutions (www.myassays.com ). Cytokine levels in protein lysates were normalized on total protein concentrations as determined by Bradford protein assay (Biorad) and expressed as picogram of cytokine per microgram of total protein. Intracellular NAD levels were measured using the colorimetric assay NAD/NADH Quantitation assay (Sigma-Aldrich) according to the manufacturer’s protocol and expressed as fold over control.
9
Cytokine Production Measurement by ELISA
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Cytokines production were measured in supernatants of the cultured cells using ELISA kits for TNF-α, IL-1β, IFN-α, and IL6 (R&D Systems, Minneapolis, MN) following the instructions of the manufacturer.
10
Inflammation and Oxidative Stress in ePE
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