Luminescence based assay

Manufactured by Promega

Sourced in United States

Luminescence-based assays are a type of analytical technique used to detect and quantify the presence of specific molecules or analytes in a sample. The assay utilizes a luminescent reaction to generate light, which can be measured and correlated to the concentration of the target analyte. This provides a sensitive and quantitative method for various applications in biological research and clinical diagnostics.

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Lab products found in correlation

Oxidative dna damage elisa kit, by Cell Biolabs (1 mentions) Firefly luciferase assay kit, by Promega (1 mentions) Celltiter glo reagent, by Promega (1 mentions)

2 protocols using luminescence based assay

Quantification of Cellular Redox Status

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A total of 24 h post treatments, we washed the spheroids twice with PBS and incubated them in 50 µL TrypLE express reagent (12604-021, Life Technologies Europe B.V., Bleiswijk, The Netherlands) for 15 min at 37 °C in a 96-well plate. Immediately after, we added fresh warm culture medium and we dissociated the spheroids with gentle mechanical force using a Pasteur pipette. After the cells were centrifuged at 1000 rpm up to 3 min, we measured the glutathione (GSH) and 8-hydroxy-2′-deoxyguanosine (8-OHdG) levels. We quantified the total GSH and genomic DNA in 10⁴ cells from dissociated spheroids. We measured the GSH level using a luminescence-based assay (Promega, Madison, WI, USA) following a standard molecular biology protocol [37 (link),38 (link)]. We used the genomic DNA extracted from the same number of tumor cells for the detection of the 8-OHdG level using an oxidative DNA damage ELISA kit (Cell Biolabs, inc. San Diego, CA, USA).

Shaw P., Kumar N., Privat-Maldonado A., Smits E, & Bogaerts A. (2021). Cold Atmospheric Plasma Increases Temozolomide Sensitivity of Three-Dimensional Glioblastoma Spheroids via Oxidative Stress-Mediated DNA Damage. Cancers, 13(8), 1780.

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Proliferation, Redox, and CRISPR Assays

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For all relevant proliferation assays, we used CellTiter-Glo reagent (Promega) as per manufacturer’s protocol. Luciferase assays were performed using Firefly luciferase assay kit as per manufacturer’s protocol (Promega). Luciferase assay values were either normalized to total protein content measured by Bradford assay or viable cells measured by CellTiter-Glo. Reduced and oxidized glutathione levels were measured using luminescence-based assay (Promega, V6611) as per manufacturer’s instructions. pNLF1-NRF2 (NRF2-nanoluc fusion, Promega) was used to assess NRF2 stability. For each biological replicate relevant reporter assays were done in duplicate.
To detect CRISPR-induced mutations at target loci we first harvested genomic DNA (Promega), amplified the target region, and performed T7 assay (NEB) as per manufacturer’s protocol.

Sanghvi V.R., Leibold J., Mina M., Mohan P., Berishaj M., Li Z., Miele M.M., Lailler N., Zhao C., de Stanchina E., Viale A., Akkari L., Lowe S.W., Ciriello G., Hendrickson R.C, & Wendel H.G. (2019). The oncogenic action of NRF2 depends on de-glycation by Fructosamine-3-kinase. Cell, 178(4), 807-819.e21.

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