Please use this identifier to cite or link to this item:
https://hdl.handle.net/10316/101007
DC Field | Value | Language |
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dc.contributor.author | Grilo, Luís F | - |
dc.contributor.author | Martins, João D | - |
dc.contributor.author | Cavallaro, Chiara H | - |
dc.contributor.author | Nathanielsz, Peter W | - |
dc.contributor.author | Oliveira, Paulo J | - |
dc.contributor.author | Pereira, Susana P | - |
dc.date.accessioned | 2022-07-25T23:01:31Z | - |
dc.date.available | 2022-07-25T23:01:31Z | - |
dc.date.issued | 2020-12 | - |
dc.identifier.issn | 08872333 | pt |
dc.identifier.uri | https://hdl.handle.net/10316/101007 | - |
dc.description.abstract | Oxidative stress biomarkers are powerful endpoints in toxicological research. Cellular reductive/oxidative balance affects numerous signaling pathways involving H2O2. Detoxification and control of H2O2 levels results mainly from catalase activity. The aim of this work was to develop a precise, simple, cost-effective microassay to measure catalase activity in small tissue samples and cell extracts. We developed a protocol that quantifies H2O2 decomposition by intrinsic catalase in biological samples. Catalase activity was calculated based on rate of decomposition of H2O2, following absorbance at 240 nm. We developed a multi-well spectroscopic approach, reducing sample quantity requirements and allowing simultaneous assessment of large number of samples. The protocol is sensitive across a wide range of catalase activity (11.5-7575 U). The assay presents a 95% confidence interval with an intra-assay coefficient of variation of 3.7%, an inter-assay coefficient of variation of 6.2% and good correlation with a commercial kit. The assay was established and validated for different biological samples, including sheep hepatic tissue and human tumor and non-tumor cell lines. This high-throughput method is robust, sensitive, time-saving and cost-effective, generating highly reproducible results with precision and good correlation with a commercial kit reinforcing the method's validity for research and toxicological applications. | pt |
dc.language.iso | por | pt |
dc.rights | openAccess | pt |
dc.subject | Antioxidant defense; Catalase; Hydrogen peroxide; Oxidative stress; Toxicity screening | pt |
dc.subject.mesh | Animals | pt |
dc.subject.mesh | Catalase | pt |
dc.subject.mesh | Hep G2 Cells | pt |
dc.subject.mesh | Humans | pt |
dc.subject.mesh | Hydrogen Peroxide | pt |
dc.subject.mesh | Kinetics | pt |
dc.subject.mesh | Liver | pt |
dc.subject.mesh | Rotenone | pt |
dc.subject.mesh | Sheep | pt |
dc.subject.mesh | Vitamin K 3 | pt |
dc.subject.mesh | tert-Butylhydroperoxide | pt |
dc.subject.mesh | Biological Assay | pt |
dc.subject.mesh | High-Throughput Screening Assays | pt |
dc.title | Development of a 96-well based assay for kinetic determination of catalase enzymatic-activity in biological samples | pt |
dc.type | article | - |
degois.publication.firstPage | 104996 | pt |
dc.peerreviewed | yes | pt |
dc.identifier.doi | 10.1016/j.tiv.2020.104996 | pt |
degois.publication.volume | 69 | pt |
dc.date.embargo | 2020-12-01 | * |
uc.date.periodoEmbargo | 0 | pt |
item.fulltext | Com Texto completo | - |
item.grantfulltext | open | - |
item.languageiso639-1 | pt | - |
item.cerifentitytype | Publications | - |
item.openairetype | article | - |
item.openairecristype | http://purl.org/coar/resource_type/c_18cf | - |
Appears in Collections: | I&D CNC - Artigos em Revistas Internacionais |
Files in This Item:
File | Description | Size | Format | |
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Grilo et al. - 2020 - Toxicology in Vitro-annotated.pdf | 1.55 MB | Adobe PDF | View/Open |
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