To analyze TNF-α release, isolated PBMC (2 × 106 cells/mL) were pretreated with aqueous plant extracts in the concentration of 1–333 μg/mL for 3 h or solvent control (10% water) for 3 h and subsequently stimulated with 1 μg/mL LPS for 3 h at 37°C in a humidified incubator with 5% CO2/95% air atmosphere. Supernatants were then used for photometric quantification of TNF-α using the TNF-α ELISA kit supplied from eBioscience (Frankfurt, Germany) according to the manufacturer's instructions.
Tnf α elisa kit
Manufactured by Thermo Fisher Scientific
Sourced in United States, China, Austria
The TNF-α ELISA kit is a quantitative immunoassay designed to measure the concentration of tumor necrosis factor alpha (TNF-α) in biological samples. It utilizes the enzyme-linked immunosorbent assay (ELISA) technique to detect and quantify the target analyte.
Automatically generated - may contain errors
Lab products found in correlation
Il 6 elisa kit, by Thermo Fisher Scientific (13 mentions)
Il 1β elisa kit, by Thermo Fisher Scientific (8 mentions)
Lipopolysaccharides (lps), by Merck Group (4 mentions)
Interleukin 6 (il 6), by Thermo Fisher Scientific (3 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Oil red o, by Merck Group (2 mentions)
Phorbol myristate acetate (pma), by Merck Group (2 mentions)
Human il 6 elisa kit, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Eclipse ci microscope, by Nikon (2 mentions)
Chemray 240, by Rayto (2 mentions)
Chemray 800, by Rayto (2 mentions)
Il 10, by Thermo Fisher Scientific (2 mentions)
Ifn γ, by Thermo Fisher Scientific (2 mentions)
Il 4 elisa kit, by Thermo Fisher Scientific (1 mentions)
Mouse ifn γ elisa kit, by Thermo Fisher Scientific (1 mentions)
Human ifn γ elisa ready set go kit, by Thermo Fisher Scientific (1 mentions)
Fluo 3 am, by Thermo Fisher Scientific (1 mentions)
Rifampicin, by Merck Group (1 mentions)
Nahco3, by Merck Group (1 mentions)
97 protocols using tnf α elisa kit
1
Quantifying TNF-α Release in PBMC
2
Serum Biomarker Analysis by ELISA
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Peripheral blood was collected. Serum was separated by centrifugation for 10-15 min at 3000 rpm and stored at −80°C for analysis. The secretion of sRAGE, TNF-α, IL-1β, IL-6, TGF-β1, and ox-LDL in serum was detected by ELISA assay (sRAGE, IL-6, IL-1β, and TGF-β1 ELISA kit, BOSTE, Wuhan, Hubei; TNF-α ELISA kit, eBioscience, San Diego, CA; ox-LDL ELISA kit, Nanjing Xinfan Biology, Nanjing, China). All measurements were carried out strictly according to the manufacturer's instructions.
3
Cytokine Secretion Assay for Splenocytes
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Single cell suspensions were acquired and diluted in 2 ml culture medium containing the same concentration of stimulus as described above to the 12-well plates at 1×106 cells per well. The plates were incubated at 37°C, 5% CO2, 100% humidity for 36 h. The suspensions were collected of the splenocyte cultures for sandwich ELISA to detect the level of the cytokines. The cell deposits were harvested to prepare for flow cytometry analysis. We used the mouse IFN-γ ELISA kit, TNF-α ELISA kit and IL-4 ELISA kit (eBioscience, Inc., San Diego, CA, USA). The concentration of the cytokines was calculated in the suspension according to the standards curve.
4
Compound 1 Modulates Inflammatory Cytokines
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RAW 264.7 cells were plated in 24-well plates at a density of 4 × 105 cells/well and incubated for 24 h at 37 °C. Cells were then exposed to compound 1 at concentrations of 50 and 100 μM for 1 h at 37 °C prior to incubation with 1 μg/mL LPS for 24 h at 37 °C. To determine IL-6 and tumor necrosis factor alpha (TNF-α) production, the supernatant was obtained and assayed to quantify the levels of these cytokines using an IL-6 sandwich enzyme-linked immunosorbent assay (ELISA) kit (BD Biosciences, CA, USA) and a TNF-α ELISA kit (eBiosciences, CA, USA), according to the manufacturer’s instructions.
5
Quantification of IFN-γ and TNF-α
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To analyze for release of IFN-γ or TNF-α, s upernatants were used for photometric quantification by the human IFN-γ ELISA Ready-Set-Go kit and the TNF-α ELISA kit, respectively, both supplied from eBioscience (Frankfurt, Germany) according to the manufacturer’s instructions.
6
Mast Cell Activation Assay
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Rifampicin, compound 48/80, disodium cromoglycate (cromolyn), p-nitrophenyl-N-acetyl-β-D-glucosaminide [PN-(GlcNAc)2], thiazolyl blue tetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), toluidine blue dye, CaCl2, MgCl2, NaHCO3, and glucose were purchased from Sigma-Aldrich Korea (Yongin, Korea). The histamine enzyme-linked immunosorbent assay (ELISA) kit was obtained from IBL International GmbH (Hamburg, Germany). We purchased the human PGD2 ELISA kit from Cusabio (Wuhan, China) and the TNF-α ELISA kit from eBioscience (San Diego, USA). The total RNA isolation kit was purchased from GeneAll (Seoul, Korea). Fluo-3 AM was obtained from Molecular Probes (Eugene, OR, USA).
7
Cytokine Quantification by ELISA
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On the basis of the instructions, IL-6 and TNF-α levels were verified using IL-6 ELISA kit (eBioscience, CA, USA) and TNF-α ELISA kit (eBioscience, CA, USA), respectively [27 (link)].
8
Quantifying Colonic TNF-α Levels
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Fifty milligrams of tissue from the distal portion of the colon were washed with 1x PBS and then cut into segments of ≈ 1 cm2. Colonic tissue samples were cultured in RPMI 1640 containing 5% fetal bovine serum in a 24-well culture plate for 24 h (24 (link)). Supernatants were collected for the detection of sTNF-α; and membrane proteins were extracted according to the manufacturer’s protocol (Biovision, Milpitas, CA, USA) for the detection of tmTNF-α. Concentrations of both forms of TNF-α in colonic tissue or serum sTNF-α were detected using a TNF-α ELISA kit (eBioscience, San Diego, CA, USA). The ratio of tmTNF-α and sTNF-α in colonic tissue was calculated.
9
Quantifying Inflammatory Cytokines and MMPs
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Commercially available ELISA kits used for measuring inflammatory cytokines and MMPs were as follows: IL-6, IL-8, and IFN-γ ELISA kits from Neobioscience Technology Co, Ltd (Beijing, China); IL-17 ELISA kit from QuantoBio Biotechnology Co, Ltd (Beijing, China); TNF-α ELISA kit from eBioscience; and MMP-3 ELISA kit from R & D Systems (Minneapolis, Minnesota, USA).
10
Serum TNF-α Quantification by ELISA
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Serum TNF‐α level was analysed by ELISA per manufacturer's instructions (TNF‐α ELISA kit: eBioscience Cat# BMS607/3 RRID:AB2575663).
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