BV-2 cells were treated for 24 h and then supernatants from controls (dH2O only), from cells treated with GLE (0.5 mg/ml) only, LPS only (1.0 μg/ml), and GLE (0.5 mg/ml) + LPS (1.0 μg/ml) (where LPS was added after 1 h incubation with GLE), were used in the assay. Specific ELISAs (RayBiotech, Norcross, GA, USA) were performed using G-CSF (Cat# ELM-G-CSF), IL1α (Cat# ELM-IL1a), MCP-5 (Cat# ELM-MCP5), MIP3α (Cat# ELM-MIP3a), and RANTES (Cat# ELM-RANTES) following manufacturer’s instructions and as previously described (Mendonca et al., 2017 (link)). Briefly, 100 μl of supernatant from samples and standards were added to 96 well plates pre-coated with the capture antibody. After incubation, 100 μl of prepared biotinylated antibody mixture was added to each well and incubated for 1 h. The mixture was then decanted, and streptavidin solution (100 μl) was placed in each well and incubated. Substrate reagent (100 μl) was then added to each well for 30 min, followed by the addition of stop solution (50 μl). Data were quantified by optical density at 450 nm.
Specific elisas
Manufactured by RayBiotech
Sourced in United States
Specific ELISAs are quantitative immunoassay kits designed to detect and measure the concentration of a particular target analyte in a sample. The core function is to provide a reliable and sensitive method for the quantitative analysis of the target molecule.
Automatically generated - may contain errors
2 protocols using specific elisas
1
ELISA Analysis of Inflammatory Factors in BV-2 Cells
2
ELISA Analysis of Inflammatory Factors in BV-2 Cells
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BV-2 cells were treated for 24 h and then supernatants from controls (dH2O only), from cells treated with GLE (0.5 mg/ml) only, LPS only (1.0 μg/ml), and GLE (0.5 mg/ml) + LPS (1.0 μg/ml) (where LPS was added after 1 h incubation with GLE), were used in the assay. Specific ELISAs (RayBiotech, Norcross, GA, USA) were performed using G-CSF (Cat# ELM-G-CSF), IL1α (Cat# ELM-IL1a), MCP-5 (Cat# ELM-MCP5), MIP3α (Cat# ELM-MIP3a), and RANTES (Cat# ELM-RANTES) following manufacturer’s instructions and as previously described (Mendonca et al., 2017 (link)). Briefly, 100 μl of supernatant from samples and standards were added to 96 well plates pre-coated with the capture antibody. After incubation, 100 μl of prepared biotinylated antibody mixture was added to each well and incubated for 1 h. The mixture was then decanted, and streptavidin solution (100 μl) was placed in each well and incubated. Substrate reagent (100 μl) was then added to each well for 30 min, followed by the addition of stop solution (50 μl). Data were quantified by optical density at 450 nm.
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