Cell culture medium was collected after treatments, and HA levels were determined by using the HA Test Kit (Corgenix, Broomfield, CO, USA). IL-10 levels in skin and wounds were measured in tissue homogenate by using the Quantikine IL-10 ELISA Kit (R&D Systems, Minneapolis, MN, USA). Data were normalized against total protein in each sample and calculated by using the Coomassie Plus protein assay (Thermo Fisher Scientific).
Il 10 quantikine elisa kit
Manufactured by R&D Systems
Sourced in United States
The IL-10 Quantikine ELISA Kit is a quantitative sandwich enzyme immunoassay designed to measure human interleukin-10 levels in cell culture supernatants, serum, and plasma. It utilizes a microplate pre-coated with an antibody specific for IL-10.
Automatically generated - may contain errors
Lab products found in correlation
Coomassie plus protein assay, by Thermo Fisher Scientific (1 mentions)
Infinite 200 pro nanoquant, by Tecan (1 mentions)
Complete mini edta free protease inhibitor cocktail, by Merck Group (1 mentions)
T per tissue protein extraction reagent, by Thermo Fisher Scientific (1 mentions)
Tnf α quantikine elisa kit, by R&D Systems (1 mentions)
Il 1β quantikine elisa kit, by R&D Systems (1 mentions)
Il 6 quantikine elisa kit, by R&D Systems (1 mentions)
Microplate reader, by Bio-Rad (1 mentions)
T per reagent, by Thermo Fisher Scientific (1 mentions)
Protease inhibitor, by Roche (1 mentions)
6 protocols using il 10 quantikine elisa kit
1
Quantification of HA and IL-10 Levels
2
Quantification of Tumor-Derived IL-10 and IgG4 Levels
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For the IL-10 ELISA, tumor cells were co-cultured with B cells for 24-h. Control cells were incubated with media only. To ensure detection of tumor cell IL-10, B lymphocytes and media were removed, and fresh media was added back to wells for a 24-h period. Supernatants were then collected, spun down to remove cell debris, and analyzed using the Quantikine IL-10 Elisa kit (R&D systems) according to manufacturer’s instructions. For the IgG4 Elisa, EBV B cells were activated and co-cultured with or without TNBC cell lines for 5 days as previously described. B cells were then removed and allowed to expand for two weeks. Cells were then plated for 24 h and supernatant was captured. Supernatants were analyzed using the Human IgG4 Elisa kit (Invitrogen). Absorbance was read at 450 nm on the Infinite® 200 PRO NanoQuant (Tecan Life Sciences). Experiments were performed in duplicate in each cell line.
3
Quantifying IL-10 in Ileum Tissue
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Ileum tissue samples weighing 100 mg were homogenized in 400 μL of T-PER™ tissue protein extraction reagent (ThermoFisher Scientific, USA) supplied with cOmplete™ mini EDTA-free protease inhibitor cocktail (Merck, Germany) using a hand-held homogenizer (T10 Basic, IKA®, China). Lysates were centrifuged at 10,000 × g for 20 min, and the resulting supernatants were analyzed with IL-10 Quantikine ELISA Kit (R&D systems, USA) to measure IL-10 concentration.
4
Quantitative Cytokine Profiling by ELISA
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The level of each protein, TGF-β, IL-1β, IL-10, IL-6 and TNF-α, was determined with ELISA, using the following commercial kits: TGF-β Quantikine ELISA kit (cat. nο. MB100B; R&D Systems, Inc., Minneapolis, MN, USA), IL-1β Quantikine ELISA kit (cat. nο. MLB00C; R&D Systems, Inc.), IL-10 Quantikine ELISA kit (cat. nο. M1000B; R&D Systems, Inc.), IL-6 Quantikine ELISA kit (cat. nο. M6000B; R&D Systems, Inc.) and TNF-α Quantikine ELISA kit (cat. nο. MB100B; R&D Systems, Inc.) Briefly, 100 µl of each standard and sample were incubated with pre-coated antibody plates for 2.5 h at RT with gentle agitation. Following an extensive wash with the provided buffer, 100 µl of biotinylated antibody were added to the wells for 1 h at RT with gentle shaking. The amount of cytokine present in the sample or standard solution present in each well was then determined by incubating the previous mix with 100 µl of streptavidin solution for 45 min at RT and then with 100 µl of TMB One-Step substrate reagent. The colorimetric reaction was stopped after 30 min of incubation at RT by adding 50 µl of stop solution to each well. The absorbance at 450 nm was immediately measured using a microplate reader (Bio-Rad, Hercules, CA, USA).
5
Quantifying Tissue IL-10 Post-ICH
6
Enhancement of IL-10 Production in Macrophages
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Using the same experimental procedure described previously in the present study with some modification, IL-10 production enhancement in macrophages by the lysate of L. plantarum strains was examined, and 1 μg/mL LPS and 100 μg/mL of L. plantarum lysate co-treated groups were added in this experiment. After treatment with each substance, cells were incubated for another 24 h, and culture supernatant samples were collected. IL-10 concentrations of each culture supernatant of the experimental groups were determined with an IL-10 Quantikine ELISA Kit (R&D Systems, United States). The dose of lysate (100 μg/mL) was selected as a representative dose based on the previous experiment of IL-6 and TNF-α quantification.
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