Gallic acid, ellagic acid, piperine, β-asarone (Merck, Darmstadt, Germany); plumbagin (Sigma-Aldrich, Seelze, Germany); acetonitrile and phosphoric acid (Labscan, Bangkok, Thailand); purified water was prepared by Milli Q® system from Millipore (Bedford, MA, USA); murine macrophage leukemia cell line (RAW 264.7: ATCC® TIB-71™; American Type Culture Collection, VA, USA); Roswell Park Memorial Institute medium 1640 (RPMI-1640), fetal bovine serum (FBS), penicillin-streptomycin (P/S), and 0.5% trypsin-EDTA (Gibco BRL Life Technologies, NY, USA); phosphate buffer saline (PBS; Amresco, Ohio, USA); dimethyl sulfoxide (DMSO; Fluka, Munich, Germany); LPS and 3-(4,5-dimethyl-2-thiazolyl)-2,5 diphenyl-2H-tetrazolium bromide (MTT) (Sigma-Aldrich Inc., MO, USA); PGE2 EIA kit (monoclonal; Cayman Chemical Company, MI, USA); mouse TNF-α Quantikine ELISA Kit (R&D Systems Inc., MN, USA).
Mouse tnf α quantikine elisa kit
Manufactured by R&D Systems
Sourced in United States, Japan
The Mouse TNF-α Quantikine ELISA Kit is a quantitative sandwich enzyme immunoassay designed to measure mouse tumor necrosis factor alpha (TNF-α) levels in cell culture supernates, serum, and plasma.
Automatically generated - may contain errors
Lab products found in correlation
Mouse il 6 quantikine elisa kit, by R&D Systems (11 mentions)
Mouse il 10 quantikine elisa kit, by R&D Systems (2 mentions)
Il 10 mouse elisa kit, by Thermo Fisher Scientific (2 mentions)
Mouse il 6 elisa kit, by Thermo Fisher Scientific (2 mentions)
Red blood cell lysing buffer hybri max, by Merck Group (2 mentions)
Lipopolysaccharide lps, by Merck Group (2 mentions)
Il 10, by Thermo Fisher Scientific (2 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Mouse il 1β quantikine elisa kit, by R&D Systems (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Ellagic acid, by Merck Group (1 mentions)
Piperine, by Merck Group (1 mentions)
Plumbagin, by Merck Group (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Pge2 eia kit, by Cayman Chemical (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Milli q system, by Merck Group (1 mentions)
Gallic acid, by Merck Group (1 mentions)
Raw 264.7 atcc tib 71, by American Type Culture Collection (1 mentions)
Mouse lipocalin 2 ngal quantikine elisa kit, by R&D Systems (1 mentions)
39 protocols using mouse tnf α quantikine elisa kit
1
Anti-inflammatory potentials of natural compounds
2
Multiplex Biomarker Quantification
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The plasma and urine levels of mediators and biomarkers (IL-10, TNF-α, IL-6, CXCL1, CXCL2, HMGB1, IGFBP7, TIMP-2, NGAL, L-FABP, and PCX) were analyzed using commercial available ELISA kits (Mouse IL-10 Quantikine ELISA Kit, R&D Systems, M1000B; Mouse TNFα Quantikine ELISA Kit, R&D Systems, MTA00B; Mouse IL-6 Quantikine ELISA Kit, R&D Systems, M6000B; Mouse CXCL1/KC Quantikine ELISA Kit, R&D Systems, MKC00B; Mouse CXCL2/MIP2 Quantikine ELISA Kit, R&D Systems, MM200; HMGB1 ELISA, IBL-International, ST51011; IGFBP7 ELISA Kit, Antikörper-Online, ABIN816449, Mouse TIMP-2 DuoSet ELISA, R&D Systems, DY6304-05, Mouse Lipocalin-2/NGAL Quantikine ELISA Kit, R&D Systems, MLCN20, Mouse/Rat FABP1/L-FABP Quantikine ELISA Kit, R&D Systems, RFBP10: PODXL ELISA Kit Mouse, Aviva Systems Biology, OKEH03213). The ELISA plates were quantitatively analyzed on a BioTek Synergy 2 plate reader.
3
Evaluating TNF-α Production and Cytotoxicity
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RAW264.7 cells were seeded into cell culture plates. For immunofluorescence imaging, coverslips were plated and the cells were seeded onto coverslips. The plasmid constructs were transfected into cells using PEI-Max (Polysciences, Inc., Warrington, PA, United States) according to the manufacturer’s instructions. To examine TNF-α production by the transfected cells, cell culture supernatants were collected 24 h after transfection. TNF-α levels in the culture supernatants were measured using a mouse TNF-α Quantikine ELISA Kit (R&D Systems, Minneapolis, MN, United States) according to the manufacturer’s instructions. For treatment with recombinant proteins, 10 μg/ml or the indicated concentrations of recombinant proteins purified from E. coli were added to culture supernatants for the indicated durations. Control green fluorescent protein (GFP) were boiled to denature state before treatment for avoiding unexpected effect. Lactate dehydrogenase (LDH) release in culture supernatants were measured using a LDH Cytotoxicity Assay Kit (Nacalai) according to the manufacturer’s instructions.
4
Quantifying Mouse TNF-α via ELISA
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The quantity of TNF-α was measured using a Mouse TNF-α Quantikine ELISA kit purchased from R&D systems (Minneapolis, MN, USA). After pretreatment with each herbal extract for 2 h, BV-2 cells were then stimulated with LPS (1 μ/mL) for an additional 22 h. Cell culture supernatants were transferred to 96-well microplates coated with mouse anti-TNF-α antibody and then incubated with TNF-α conjugate. The concentration of TNF-α in each well was calculated by comparison with the standard concentrations provided in the kit according to manufacturer instructions.
5
Quantification of TNF-α and OPN Proteins
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The concentration of TNF‐α and OPN protein in cell‐free culture supernatants was determined by solid‐phase ELISA using the Mouse TNF‐α Quantikine ELISA Kit (R&D Systems) and Horse OPN ELISA Kit (MyBioSource, San Diego, CA, USA), respectively. Procedure and calculations were carried out in accordance with the manufacturer's instruction. The absorbance was measured with a 96‐well microplate reader (Epoch; BioTek) at 450 nm.
6
Quantification of Mouse TNF-α
7
Quantitative Cytokine and Protein Analysis
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The concentration of TNF-α and p53 protein in cell-free culture supernatants was determined by solid phase ELISA using the Mouse TNF-α Quantikine ELISA Kit (R&D Systems) and Horse p53 ELISA Kit (MyBioSource, San Diego, CA, USA), respectively. All steps in the procedure and calculations were carried out according to the manufacturer’s instruction. The absorbance was measured with a 96-well microplate reader (Spectrostar Nano, BMG Labtech) at 450 nm.
8
Quantifying Plasma TNF-α Levels
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The concentration of TNF-α in plasma was evaluated by Mouse TNF-α Quantikine ELISA kit according to manufacturer instructions (R&D Systems, Minneapolis, MN, USA).
9
Quantification of IL-6 and TNF-α
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IL-6 Quantikine ELISA Kit (M6000B; R&D Systems, Minneapolis, MN, USA) and mouse TNF-α Quantikine ELISA Kit (MTA00B; R&D Systems) were used to detect the expression of IL-6 and TNF-α in serum, respectively. The test steps were operated according to the instructions of the kit.
The absorbance value was obtained at the wavelength of 450nm using enzyme-labeling instrument, and the standard curve was used to calculate the corresponding concentration value.
The absorbance value was obtained at the wavelength of 450nm using enzyme-labeling instrument, and the standard curve was used to calculate the corresponding concentration value.
10
Regulation of Monocyte TNF-α Secretion
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Mouse monocytes (0.5 × 106/ml) were plated in 24-well plates and starved overnight by growing in minimal essential medium containing 0.5% fetal bovine serum and antibiotics. On the following day, the plates were replaced with fresh starving medium and treated with increasing concentrations of HNK, MCP, and MCP : HNK (9 : 1) in the presence and absence of LPS. The compound was added initially and after incubation for 2 h at 37°C; 20 ng/ml LPS was added to induce an inflammatory response. The plate was incubated for an additional 4 h and culture medium was collected, centrifuged, and stored at −80°C. TNF-α produced and secreted into the medium by the cells was analyzed by ELISA protocol using the mouse TNF-α Quantikine ELISA kit (R&D systems, Minneapolis, MN) as per manufacturer's instructions [43 (link)].
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