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Oxidative Stress

Aug 21, 2017

Oxidative stress reflects the relationship between production of reactive oxidative species (ROS) and the ability of a biological system to cope with ROS.  Excessive oxidative stress damages cellular components, such as lipids, proteins, and DNA, and ultimately can lead to cell death (Figure 1).  In vivo, oxidative stress is a precursor to a number to a number of toxicological effects, including skin and lung sensitization and carcinogenicity.  As such, evaluating this mode of action is an important step in elucidating the potential risk from a chemical. 

ScitoVation has a great deal of experience in quantitative evaluation of oxidative stress and cellular response, including expertise in the application of following assays and kits:

 

ROS detection:

Detection of intracellular ROS - H2DCF-DA probe with Flow Cytometry detection

roGFP Glutathione redox potential detection assay – live cell imaging with EGFP reporter in HaCaT cell line

CellROX Deep Red Reagent for oxidative stress detection – Flow cytometry and high content imaging detection

 

Cellular damage:

Protein oxidative damage (protein carbonylation, nitration) - ELISA

Lipid peroxidation - C11-BODIPY581/591 Image-iT Kit with flow cytometry detection

DNA/RNA oxidative damage (8-OHdG) – ELISA

Cytotoxicity and apoptosis - ApoTox-Glo Triplex plate reader assay

Cytotoxicity using readouts for LDH release, nuclear count, membrane permeability, nuclear morphology, ATP, MTT, Caspase 3/7, etc. – detection via plate reader, flow cytometry, or high content imaging

 

Antioxidant response:

Total antioxidant capacity assay – ELISA

GSH depletion - GSH-Glo Glutathione assay

 

Nrf2 accumulation – Flow cytometry detection

Nrf2 nuclear accumulation/translocation – High content imaging based detection

Nrf2 nuclear accumulation/translocation -  Quantitative Western blot analysis (ProteinSimple)

ARE luciferase reporter assay

Antioxidant gene expression (NQO1, HO1, etc.) - qRT-PCR detection

Transcriptomic gene signature analysis – microarray and RNA-Seq

 

Contact us for further information!