LDH release was measured using CytoTox 96 Non-Radioactive Cytotoxicity Assay (Promega) according to manufacturer’s instructions. Data calculated as percent death (signal over max death after treatment with 0.1% Triton). YOYO-1 (491/509) dye (at 200 nM final concentration) was added at the time of stimulation and scanned in green channel every 30 min to 1 h for at least 16 h on the Essen BioScience IncuCyte ZOOM at 10× magnification. Nuclear-ID was added at the last time point and scanned in red channel. IncuCyte software was used to determine total number of dead YOYO+ cells and Nuclear-ID+ (live and dead). Percent death (or percent YOYO+) was calculated as the number of YOYO+ cells divided by the total number of NuclearID+ cells. IL-1β was measured from cell culture supernatants and serum by mouse IL-1β tissue culture kit (Meso Scale Discovery). IL-18 was measured from cell culture supernatants and serum using mouse IL-18 ELISA (MBL International).
Mouse il 1β tissue culture kit
Manufactured by Mesoscale
Sourced in United States
The Mouse IL-1β tissue culture kit is a laboratory tool designed for the quantitative measurement of mouse interleukin-1 beta (IL-1β) levels in cell culture supernatants. It provides the necessary reagents and protocols to perform this analysis.
Automatically generated - may contain errors
3 protocols using mouse il 1β tissue culture kit
1
Cytotoxicity and Inflammatory Markers Assay
2
Cytotoxicity Assay and Live-Cell Imaging
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LDH release was measured using CytoTox 96 Non-Radioactive Cytotoxicity Assay (Promega) according to manufacturer’s instructions. Data calculated as signal over max death signal (cells treated with 0.1% Triton). YOYO-1 (491/509) dye (at 200 nM final concentration) was added at the time of stimulation and scanned every 30 min–1 hr for at least 16 hours on the Essen BioScience IncuCyte ZOOM at 10× magnification, scanned in green channel. Nuclear-ID was added at the last time point, scanned in red channel. IncuCyte software was used to determine total number of dead YOYO+ cells and Nuclear-ID+ (live and dead). Percent death (or % YOYO+) was calculated as the number of YOYO+ cells divided by the total number of NuclearID+ positive cells. IL-1β was measured from cell culture supernatants by mouse IL-1β Tissue culture kit (Meso Scale Discovery).
3
Cytokine Quantification in Biofluids
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Cytokine levels in culture medium and serum were determined by magnetic-bead–based multiplex assay using Luminex technology (Bio-Rad, Hercules, CA, USA), IL-1β ELISA (R&D Systems, Minneapolis, MN, USA) and mouse IL-1β tissue culture kit (Meso Scale Discovery, Rockville, MD, USA) according to the manufacturers’ instructions.
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