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Safire 2 microplate luminometer

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The Safire II Microplate Luminometer is a compact and versatile instrument designed for sensitive luminescence detection in microplate assays. It offers high-performance photon counting capabilities for a wide range of applications, including cell-based assays, reporter gene analyses, and luminescent enzyme activity measurements.

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Safire (53 citations) Microplate luminometer (6 citations)

2 protocols using safire 2 microplate luminometer

1

Quantification of O6-methylguanine DNA Adducts

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Exposure to certain NOCs causes O6‐methylguanine (O6‐MeG) adducts formation. O6‐MeG DNA adducts are also mutagenic and have frequently been shown to occur in human CRC tissue.[51, 52] Samples were analyzed using an ELISA method described by Georgiadis et al.[53] In short, restriction enzymes were used to digest DNA to fragments of size expected to contain no more than one O6‐meG residue. Anti‐adduct antisera were used to transfer O6‐meG–containing fragments to a solid surface, where they were detected using anti‐ssDNA antisera. A chemiluminescence signal was finally generated by the addition of CDP‐Star substrate containing Emerald II enhancer. Chemiluminescence was measured using a Safire II Microplate Luminometer (TECAN) at 542 nm. For the quantitation of O6‐meG in unknown DNA samples, a standard curve consisting of DNA standards containing different levels of O6‐meG were prepared with DNA from HeLa cells. The assay has a limit of detection of 1.5 adducts per 109 nucleotides using 10 µg of DNA.
van Breda S.G., Mathijs K., Pieters H., Sági‐Kiss V., Kuhnle G.G., Georgiadis P., Saccani G., Parolari G., Virgili R., Sinha R., Hemke G., Hung Y., Verbeke W., Masclee A.A., Vleugels‐Simon C.B., van Bodegraven A.A, & de Kok T.M. (2021). Replacement of Nitrite in Meat Products by Natural Bioactive Compounds Results in Reduced Exposure to N‐Nitroso Compounds: The PHYTOME Project. Molecular Nutrition & Food Research, 65(20), 2001214.
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2

Quantification of O6-methylguanine DNA Adducts

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Exposure to certain NOCs causes O6-methylguanine (O6-MeG) adducts formation. O6-MeG DNA adducts are also mutagenic and have frequently been shown to occur in human CRC tissue (51 (link), 52 (link)). Samples were analysed using an ELISA method described by Georgiadis et al. (2011) (53 (link)). In short, restriction enzymes were used to digest DNA to fragments of size expected to contain no more than one O6-meG residue. Anti-adduct antisera were used to transfer O6-meG–containing fragments to a solid surface, where they were detected using anti-ssDNA antisera. A chemiluminescence signal was finally generated by the addition of CDP-Star substrate containing Emerald II enhancer. Chemiluminescence was measured using a Safire II Microplate Luminometer (TECAN) at 542 nm. For the quantitation of O6-meG in unknown DNA samples, a standard curve consisting of DNA standards containing different levels of O6-meG were prepared with DNA from HeLa cells. The assay has a limit of detection of 1.5 adducts/109 nucleotides using 10 μg of DNA.
van Breda S.G., Mathijs K., Pieters H., Sági‐Kiss V., Kuhnle G.G., Georgiadis P., Saccani G., Parolari G., Virgili R., Sinha R., Hemke G., Hung Y., Verbeke W., Masclee A.A., Vleugels‐Simon C.B., van Bodegraven A.A, & de Kok T.M. (2021). Replacement of Nitrite in Meat Products by Natural Bioactive Compounds Results in Reduced Exposure to N‐Nitroso Compounds: The PHYTOME Project. Molecular Nutrition & Food Research, 65(20), 2001214.
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