The effector cells (NTD T cells, CD19-BBz CAR, or NKG2D-BBz CAR-T cells) were co-cultured with target cells (U-251MG glioblastoma cells) for 16 h at an E:T ratio of 5:1 and the medium supernatant was assessed for the levels of cytokine secretion. The concentrations of IL-2, IL-10, TNF-α, and IFN-γ were measured using enzyme-linked immunosorbent assay kits (BD Biosciences, USA) according to the manufacturer’s instructions.
Enzyme linked immunosorbent assay kit
Manufactured by BD
Sourced in United States
Enzyme-linked immunosorbent assay (ELISA) kits are laboratory tools used to detect and quantify specific proteins, hormones, or other analytes in a sample. The kits utilize antibodies coupled with enzymes to enable sensitive and accurate measurements.
Automatically generated - may contain errors
Lab products found in correlation
Cobas integra 400 plus, by Roche (2 mentions)
Vettest 8008, by IDEXX (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Enzyme linked immunosorbent assay kit, by Thermo Fisher Scientific (1 mentions)
Lps from escherichia coli 055 b5, by Merck Group (1 mentions)
Lipopolysaccharide lps, by Merck Group (1 mentions)
Antibiotic antimycotic solution, by Merck Group (1 mentions)
Raw 264.7 macrophages, by Korean Cell Line Bank (1 mentions)
Heat inactivated fetal bovine serum, by Welgene (1 mentions)
Tbars assay kit, by Cayman Chemical (1 mentions)
Cd4 t cell isolation kit, by Miltenyi Biotec (1 mentions)
Enzyme linked immunosorbent assay kit, by Cusabio (1 mentions)
19 protocols using enzyme linked immunosorbent assay kit
1
Cytokine Secretion Assay for Effector Cells
2
Ischemia/Reperfusion Injury Biomarkers
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Blood samples (0.3 mL/rat) were taken from all rats 10 h before ischemia (baseline), after 12 h of reperfusion, and at the end of 24 h of reperfusion. However, the blood could not be collected at each hour during ischemia since blood sampling under ischemic conditions could lead to risks of complications and adverse effects such as hemolysis. Serum samples were collected by centrifugation at 3,000 rpm for 15 min and stored at −80°C until analysis. The level of IL-6, an inflammation marker, was quantified by enzyme-linked immunosorbent assay kits (BD Pharmingen, San Diego, CA, USA) according to the manufacturer’s instructions. Based on a previous study,17 (link) which showed that harmful effects caused by intestinal ischemia were not limited to the affected intestine (e.g., hepatic hypoperfusion and acute hepatic damage after I/R), serum biochemistry analysis also included AST, alanine aminotransferase (ALT), and albumin as part of a liver function test, as well as LDH as a general indicator of the presence and severity of acute or chronic tissue damage in this study. These serum levels were measured by an autoanalyzer using a commercially available test kit (VET TEST 8008; IDEXX Laboratories, Westbrook, ME, USA).
3
Quantification of IL-8 and PDGF
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Concentrations of IL-8 and PDGF in the supernatant were measured using enzyme-linked immunosorbent assay kits (BD Bioscience, Bedford, MA and Abcam Inc. Cambridge, MA, respectively).
4
Complement Activation Measurement
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Complement activation products; that is, C3a, C5a and C5b‐9, were measured in the urine and plasma samples using enzyme‐linked immunosorbent assay kits (BD Biosciences, San Diego, CA, USA) according to the manufacturer's instructions. To minimize the variations from urine collection, urinary creatinine was used for correcting the urinary C3a, C5a and C5b‐9, and their level was expressed as nanogram per milligram creatinine.
5
Cytokine Profiling in Toxoplasma-Infected Murine Cells
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C57BL/6 BMDMs were differentiated and stimulated as previously described (77 (link)). Briefly, the cells were seeded (2 × 105 per well) in 96-well plates and left to adhere overnight at 37°C in 5% CO2. Cells were infected with freshly lysed GRA833xHA, ∆gra83, and GRA83c tachyzoites at an MOI of 0.5, and supernatants (200 µL) were collected at 18 h after infection and stored at −80°C. For in vivo assays, C57BL/6 mice were euthanized after 48 h of infection, and the peritoneal cavity was washed with 1 mL of PBS, spun at 400 × g for 10 min to pellet the cells. The supernatant was collected and stored at −80°C. Spleens were also collected at 48 h, mechanically disrupted by a tissue homogenizer (Ika) in 1 mL of PBS with protease inhibitor cocktail (Roche), and spun at 10,000 × g for 10 min to pellet debris. Supernatants were collected and stored at −80°C. IL-12p40 and IFN-γ levels were determined using commercially available enzyme-linked immunosorbent assay kits (BD Biosciences) according to the manufacturer’s instructions.
6
Cytokine Profiling in Murine Serum
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Mice were anesthetized with sodium pentobarbital. Blood was collected by cardiac puncture and allowed to clot at room temperature for 1.5 hr. Each sample was centrifuged to pellet the RBCs, and serum was collected and stored at -80°C. Determination of serum concentrations of Epo, IL-6, INF-γ and TNF-α were carried out using commercially available enzyme-linked immunosorbent assay kits obtained from BD or R&D Systems, according to the manufacture’s instructions. G-CSF concentrations were measured by flow cytometry using the mouse G-CSF Flex-Set bead array.
7
Cytokine Production in Cell Lines
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HT-29 and RAW 264.7 cells were seeded into a 24-well plate at a density of 1 × 106 cells per well and cultured for 24 h at 37 °C. After removing the medium, the cells were treated with 100 μL of bacterial suspension at 5 × 108 CFU/mL along with 100 ng/mL (final concentration) LPS and incubated for 24 h at 37 °C. The culture supernatant was collected to determine interleukin (IL)-6, tumor necrosis factor (TNF)-α, and IL-8 levels using enzyme-linked immunosorbent assay kits (BD Biosciences, San Jose, CA, USA), according to the manufacturer’s instructions.
8
Quantifying Cytokine Levels in Cells
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Levels of TNFα and IL1β were quantified in cell culture supernatant using commercially available enzyme-linked immunosorbent assay kits from BD Biosciences (Franklin Lakes, NJ). enzyme-linked immunosorbent assay kits from eBioscience were used to measure IL10 and TGF-β in the cell culture supernatant.
9
Bifidobacterium Modulates Inflammatory Cytokines
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Bifidobacterium bifidum BGN4-SK and B. bifidum BGN4-pBESIL10 were adjusted to 108 CFU/mL and the HT-29 cells were treated with 100 ng/mL LPS from Escherichia coli 055: B5 (Sigma Aldrich, USA), and 100 µL neutralized cell-free supernatants (CFSs) of B. bifidum BGN4-SK, B. bifidum BGN4-pBESIL10 or a mixture of 50% (v/v) B. bifidum BGN4-SK and 50% (v/v) B. bifidum BGN4-pBESIL10, followed by incubation for 10 h at 37 ºC. Then, cell culture supernatants were collected to determine the levels of tumor necrosis factor (TNF)-α and IL-8 using enzyme-linked immunosorbent assay kits (BD Biosciences, CA, USA), according to the manufacturer’s instructions.
10
Measuring TNF-α and IL-6 in Sepsis
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The tumor necrosis factor (TNF)-α and IL-6 production in LPS-stimulated RAW264.7 cells and the serum levels of TNF-α and IL-6 were measured 6-h after CLP using commercially available enzyme-linked immunosorbent assay kits (BD Biosciences, San Jose, CA, USA) according to the manufacturer’s instructions.
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