ATP, phorbol myristate acetate, cytochalasin D and BAPTA-AM were from Sigma-Aldrich. Ultrapure lipopolysaccharide (LPS) was from Invivogen. DPI was from Calbiochem. Lipofectamine 2000 and MitoSOX were from Life Technologies. Calcium-free and calcium-containing Dulbecco's modified Eagle's medium (DMEM) were from US Biologicals. Caspase-1 inhibitor Z-YVAD-FMK and caspase-3 inhibitor Z-DEVD-FMK were from Enzo Life Sciences. The lactate dehydrogenase assay kit and in situ cell death detection kit (TUNEL) were from Roche. FLICA capase-1 kit was from Immunochemistry. Antibodies used for immunoblotting were as follows: anti-mouse caspase-1 (AG-20B-0042-C100, Adipogen, 1 μg ml−1), anti-mouse β-actin (sc-1615 horseradish peroxidase (HRP), Santa Cruz Biotechnology, 0.1 μg ml−1) and anti-mouse IL-1β (AF-401-NA, R&D Systems, 0.25 μg ml−1). Imject Alum and streptavidin-HRP were from Pierce.
Lactate dehydrogenase assay kit
Manufactured by Roche
The Lactate dehydrogenase assay kit is a laboratory tool used to measure the activity of the enzyme lactate dehydrogenase in biological samples. Lactate dehydrogenase is an important enzyme involved in energy metabolism.
Automatically generated - may contain errors
Lab products found in correlation
Multiplate reader, by Dynex (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Mitosox, by Thermo Fisher Scientific (1 mentions)
Phorbol myristate acetate (pma), by Merck Group (1 mentions)
Adenosine triphosphate (atp), by Merck Group (1 mentions)
Bapta am, by Merck Group (1 mentions)
Cytochalasin d, by Merck Group (1 mentions)
Tunel in situ cell death detection kit, by Roche (1 mentions)
Af 401 na, by R&D Systems (1 mentions)
Ultrapure lipopolysaccharide lps, by InvivoGen (1 mentions)
4 protocols using lactate dehydrogenase assay kit
1
Inflammasome Activation Assay Protocol
2
Cytotoxicity Evaluation via LDH Assay
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Cytotoxicity was evaluated using a lactate dehydrogenase assay kit (Roche, Basel, Switzerland). Briefly, 100 μL of media collected from the cell cultures on day 4 were placed in a 96-well plate and combined with 100 μL of working solution. The plate was incubated for 30 min at room temperature in the dark. The absorbance was measured at 490 nm.
3
Exosome-Activated NK Cell Cytotoxicity
4
NK Cell-Mediated Cytotoxicity Assay
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Natural killer cells (105 cells per well) were incubated with NB target cells at ratios of 1:1, 3:1, and 5:1, respectively, for 4 h in 96-well plates with the 200 µl medium per well. NK-mediated cytotoxicity was analyzed by lactate dehydrogenase assay kits (Roche, Mannheim, Germany). The procedures of the assay were followed according to the manufacturer as previously described (28 (link)). Briefly, the culture medium was collected after treatment for the assay. The reaction underwent in the dark for 30 min before measurement. Changes in the absorbance were measured at 492 nm by a multiplate reader (Dynex, UK). Results were expressed as the percentage of control. Also, evaluation of the CD107a was performed to detect the degranulation capability of NKs as described in the Section “Immunophenotyping Tests .”
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