The direct and indirect toxicity of Aβ42 oligomers was measured as lactate dehydrogenase (LDH) release, using the CytoTox® non-radioactive cytotoxicity assay kit (Promega Corporation, Madison, WI, USA) following the manufacturer’s instructions [76 (link)].
Cytotox non radioactive cytotoxicity assay kit
Manufactured by Promega
Sourced in United States
The CytoTox® non-radioactive cytotoxicity assay kit is a product offered by Promega. The kit provides a method for determining the number of cells that have died or been killed in a cell population. It measures the release of lactate dehydrogenase (LDH) from damaged cells as an indicator of cytotoxicity.
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Lab products found in correlation
6 protocols using cytotox non radioactive cytotoxicity assay kit
1
Measuring Aβ42 Oligomer Cytotoxicity
2
Candida-Induced Macrophage Cytotoxicity
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Exponential-phase Candida in PBS was diluted into serum-free, phenol red-free RPMI (GE Healthcare) and added to macrophages at an MOI of 3 for 6 h before supernatants were taken and lactate dehydrogenase release was measured using the CytoTox nonradioactive cytotoxicity assay kit (Promega).
3
Cytotoxicity Assay for DV1-Specific T Cells
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Two weeks after the third vaccination, freshly isolated mouse splenocytes were stimulated with concentrated DV1 proteins (2 μg/105 cells) for 72 h to prepare effector cells. L929 cells infected with DV1 (multiplicity of infection = 10) for 16–18 h were used as target cells. In U-bottom 96-well plates, the target cell suspension (5 × 103 cells/well) was dispensed with the effector cells at various effector: target (E:T) ratios of 100:1, 50:1, 25:1, and 5:1. After incubation for 5 h at 37°C and 5% CO2, 50 μl/well of the supernatant was transferred to 96-well flat-bottom plates. The lactate dehydrogenase (LDH) activity was determined by the CytoTox non-radioactive cytotoxicity assay kit (Promega, USA) per the manufacturer's instructions. The absorbance at 490 nm was recorded on a microplate reader, and the percentage of specific lysis (% LDH release) was calculated using the following formula: % cytotoxicity = 100 × (experimental release – effector spontaneous release – target spontaneous release)/(target maximum release – target spontaneous release).
4
MUC1-VNTR Antigen Presentation Cytotoxicity
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Mature DCs loaded with different MUC1-VNTRn antigens and autologous T-cells (ratio 1:10) were seeded into a 6-well plate supplemented with 100 U/ml IL-2 culture medium; approximately half of the medium was changed to supplement cell factors every other day. Subsequent to a 5-day culture, the cells were counted using a light microscope. A total of three groups were classified: MUC1-VNTRn group; pVAX1 without vector group; and HeLa cells group (negative control group). The organization of the groups is presented in Table II . MUC1-positive Capan2 cell strains were collected as the target cells, and the effector cells and target cells were seeded into a 96-well plate with the ratio of 40:1. The Lethal effect of CTLs was examined using the CytoTox Non-Radioactive Cytotoxicity assay kit (Promega Corporation, Madison, WI, USA), according to the manufacturer's protocol.
5
Cytotoxicity Assay for Intracellular Bacteria
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To analyze the cytotoxicity of intracellular bacteria, A549 cells were infected and treated as described for the invasion assay. However, after lysostaphin treatment, cells were washed and covered with RPMI medium supplemented with 2 mg/mL NaHCO3, 10% FBS, 1% antibiotic/antimycotic solution, and 2% MycoKill AB and incubated for up to 24 h at 37 °C and 5% CO2. Subsequently, 500 µL of culture supernatant were drawn from the well plate, centrifuged at 5000 rpm for 5 min and used to determine the bacterial cytotoxicity by measuring the activity of LDH with the CytoTox® non-radioactive cytotoxicity assay kit (Promega, Walldorf, Germany), following the manufacturer’s instructions. The higher the absorbance, the higher the LDH release from A549 cells and the bacterial cytotoxicity. As a negative control, the supernatant of A549 cells treated with 10 µL RPMI medium supplemented with 20% glycerin, and as a positive control, the supernatant of A549 cells treated with 0.09% Triton X-100, were used (the latter one applied in a 1:10 dilution).
6
Cytotoxicity Assay of CD8+ T Cells
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CD8 þ splenocytes from KLN205 tumor-bearing DBA/2 mice were isolated as above from pooled cell suspension and subsequently cocultured with 1 Â 10 4 KLN205 cells in a 96-well plate for 24 hours. Supernatant was collected and cytotoxicity was measured by CytoTox Non-Radioactive Cytotoxicity Assay kit according to the manufacturer's protocol (Promega). Cytotoxicity was calculated using the formula: effector (experimental)effector (spontaneous)target (spontaneous)/target (maximum)target (spontaneous) Â 100.
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