To challenge cells with cytokines, cells were incubated with Interferon-γ (IFNγ; Cell Signaling Technology 80385S), FasL (AdipoGen AG-40B-0130-3010), TRAIL (R&D Systems 375-TL-010), or TNF-α (AdipoGen AG-40B-0019-3010) for 24 h. TRAIL was crosslinked by incubating with anti-His Tag antibody (Thermo Fisher Scientific MA121315, 1:500) for 15 min at room temperature. Cell viability was measured using CellTiter-Glo (Promega G7571) and protein was harvested for western blots. For evaluating Caspase 8 activity, cells were incubated with FasL or crosslinked TRAIL for 3 h and harvested for Caspase 8 colorimetric assay (R&D Systems K113-100). IFNγ in the cell culture media of the T cell cytotoxic assay was quantified using an ELISA kit (Thermo Fisher Scientific KHC4021).
K113 100
Manufactured by R&D Systems
Sourced in United States
The K113-100 is a laboratory device used for quantitative analysis. The product specifications and core function are not available at this time.
Automatically generated - may contain errors
2 protocols using k113 100
1
Evaluating Cytokine-Induced Cell Responses
2
Leonurine Induces Apoptosis via Caspases
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HL-60 and U-937 cells were seeded on 6-well plates at a density of 5×10 5 cells/well (2 ml/well) and treated with leonurine at concentrations of 2, 5, and 10 μM for 48 h. Caspase-3, caspase-8, and caspase-9 activities were measured using commercial colorimetric assay kits (catalogue numbers: K106-100; K113-100; K119-100; R&D Systems, Minneapolis, MN, USA). Briefly, the control cells and leonurine-treated cells were lysed in the supplied lysis buffer. The supernatants were collected and incubated with supplied reaction buffer containing dithiothreitol and DEAD-pNA, IETD-pNA, or LEHD-pNA as substrates at 37°C for 2 h in the dark. The reactions were measured by changes in absorbance at 405 nm using a microplate reader (Thermo Fisher Scientific, Waltham, MA).
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