Peritoneal macrophages obtained from BALB/c mice and XID (2 x 105 macrophages/well) were seeded in 24-well plates. Some cultures were infected with trypomastigote form of T. cruzi overnight. After washing some cultures were stimulated with LPS (400 ng/mL) and IFN-γ (1.5 ng/mL) and co-cultivated or not with B-1 cells. Cell supernatants were collected at 24 h after stimulation for cytokine determination. IL-10, concentration was estimated by the method of sandwich immunoassay (ELISA) according to methodology recommended by the manufacturer (R&D). The optical density was evaluated by reading in a microplate spectrophotometer (Versamax Microplates Reader Molecular Devices, United States), with filter of 405 nm.
Versamax microplates reader
Manufactured by Molecular Devices
Sourced in United States
The Versamax Microplates Reader is a compact, automated instrument designed for reading and analyzing microplates. It utilizes advanced optical technology to accurately measure various types of assays, including absorbance, fluorescence, and luminescence, in a range of microplate formats.
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Lab products found in correlation
6 protocols using versamax microplates reader
1
Murine Macrophage IL-10 Modulation
2
Cytokine and PGE2 Quantification in Cell Cultures
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The concentrations of cytokines in the supernatants obtained from infected cell cultures were quantified after 24 h of incubation using the sandwich immunoassay (ELISA) method according to the methodology recommended by the manufacturer (R and D). The optical density was obtained by reading the absorbance in a plate spectrophotometer (Versamax Microplates Reader Molecular Devices, United States), with a filter of 405 nm. The concentrations of cytokines were calculated from a standard curve of recombinant cytokines. PGE2 was quantified using a PGE2-specific EIA kit, according to the methodology recommended by the manufacturer (Cayman Chemical, Ann Arbor, MI).
3
Cytokine Quantification in Macrophage-Neutrophil Co-Cultures
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The concentrations of cytokines in the supernatant obtained from co-cultures of infected macrophages and live or apoptotic neutrophils were quantified after 24 hours of incubation by the method of sandwich immunoassay (ELISA) according to methodology recommended by the manufacturer (R&D). The optical density was obtained by reading in a plate spectrophotometer (VERSAMAX MICROPLATES Reader Molecular Devices, USA). The concentrations of cytokines were calculated from a standard curve of recombinant cytokines. The determination of PGE2 (Analysis of absorption by binding enzyme immunoassay) was used EIA kit according to methodology recommended by the manufacturer (Cayman Chemical, Amn Anbor, MZ).
4
Quantification of TGF-β and TNF-α
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The concentrations of transforming growth factor beta (TGF-β) and TNF-α in the supernatant, obtained from cultures of infected macrophages cultured on 2D biofilm or 3D matrix, were quantified after 48 h of incubation by sandwich immunoassay (ELISA) according to methodology recommended by the manufacturer (R&D). The optical density was obtained from a plate spectrophotometer (VERSAMAX MICROPLATES Reader Molecular Devices, San Jose, CA, USA). The concentrations of cytokines were calculated from a standard curve of recombinant cytokines.
5
Cytokine and PGE2 Quantification
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The concentrations of cytokines in the supernatant obtained from cultures of infected B-1CDP cells or peritoneal macrophages were quantified after 24 hours of incubation by the method of sandwich immunoassay (ELISA) according to methodology recommended by the manufacturer (R&D). The optical density was obtained by reading in a plate spectrophotometer (Versamax Microplates Reader Molecular Devices, USA), with filter of 405 nm. The concentrations of cytokines were calculated from a standard curve of recombinant cytokines. Quantification of PGE2 was obtained by a PGE2 specific EIA kit, according to methodology recommended by the manufacturer (Cayman Chemical, Ann Arbor, MZ).
6
Cytokine Quantification by ELISA
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Cell supernatants were collected at 24 h postinfection for cytokine determination. TNF-α, TGF-β, and IL-10 concentration was estimated by the method of sandwich immunoassay (ELISA) according to methodology recommended by the manufacturer (R&D). The optical density was evaluated by reading in a microplate spectrophotometer (Versamax Microplates Reader Molecular Devices, USA), with filter of 405 nm.
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