Human neuroblastoma (SH-SY5Y) cells conditionally expressing human WT αSN with a Tet-Off cassette (59 (link)) were grown in RPMI supplemented with 1 μM doxycycline (to suppress αSN expression), 10% FBS, and 1% penicillin/streptomycin at 37 °C and 5% CO2. The cells were seeded in 6-well plates and differentiated for 1 day after plating using 10 μM retinoic acid in RPMI supplemented with 1% FBS and 1% penicillin/streptomycin, at 37 °C and 5% CO2 for 4 days. Media was changed every second day. Following 4 days of differentiation, cells were treated with 100 nM of the respective glycated αSN species for 4 days. The αSN species were prepared with constant shaking as described above. αSN treated the same way but without glycating agents was used as control. Cell viability was analyzed using a ToxiLight kit (Lonza) according to the manufacturer’s instructions.
Toxilight kit
Manufactured by Lonza
The ToxiLight kit is a luminescent-based assay designed to detect and quantify cytotoxicity and cell death by measuring the release of adenylate kinase (AK) from damaged cells. The kit provides a simple, rapid, and sensitive method for measuring cytotoxicity in cell-based assays.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using toxilight kit
1
Assessing Glycated α-Synuclein Cytotoxicity
2
Membrane Integrity and Cell Viability
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Cell viability in tested treatments was determined by measuring membrane integrity at the end of the exposure using ToxiLight kit (Lonza), which measures the release of cytosolic enzyme adenylate kinase into the medium. Treatments causing an increase in adenylate kinase release for more than 30% of negative control were excluded. This result corresponded well to the decrease in nuclei count of more than 50%.
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