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8 hydroxy 2 deoxyguanosine elisa kit

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The 8-hydroxy 2 deoxyguanosine ELISA Kit is a quantitative assay designed to measure the levels of 8-hydroxy-2'-deoxyguanosine, a biomarker of oxidative DNA damage, in biological samples such as urine, plasma, and cell lysates.

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ELISAs (10 citations) 8-hydroxy 2 deoxyguanosine (8-OHdG) ELISA kit (3 citations)

12 protocols using 8 hydroxy 2 deoxyguanosine elisa kit

1

Quantifying Oxidative DNA Damage

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The plasmatic levels of 8-OHdG were measured using a competitive quantitative ELISA Kit (8-hydroxy-2-deoxyguanosine ELISA Kit, Abcam), according to manufacturer instructions, in a SynergyTM multi-mode microplate reader (Gonçalves et al., 2017 (link)).
Gonçalves A.C., Alves R., Baldeiras I., Jorge J., Marques B., Paiva A., Oliveiros B., Cortesão E., Nascimento Costa J.M, & Sarmento-Ribeiro A.B. (2021). Oxidative Stress Parameters Can Predict the Response to Erythropoiesis-Stimulating Agents in Myelodysplastic Syndrome Patients. Frontiers in Cell and Developmental Biology, 9, 701328.
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2

Cellular Oxidative Stress Measurement

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2 × 105 cells with stable expression of indicated genes and/or shRNAs were seeded in a well of 6-well plate, and incubated with fresh medium overnight. Cells were treated with 12 Gy ionizing radiation, and cellular oxidative stress was measured by the following methods 3 h after irradiation. The levels of GSH/GSSG ratio were determined using the GSH/GSSG Ratio Detection Assay Kit obtained from Abcam (ab138881) and quantified using a microplate reader (Ex/Em = 490/520 nm). The level of cellular DCFDA signal was measured by using Reactive Oxygen Species (ROS) Detection Assay Kit obtained from BioVision (K936-100) and evaluated by using a microplate reader (Ex/Em = 495/529 nm). The level of 8-OHdG was examined by using 8-hydroxy 2 deoxyguanosine ELISA Kit obtained from Abcam (ab201734). The data was normalized to the cell numbers.
Hao Y., Ren T., Huang X., Li M., Lee J.H., Chen Q., Liu R, & Tang Q. (2023). Rapid phosphorylation of glucose-6-phosphate dehydrogenase by casein kinase 2 sustains redox homeostasis under ionizing radiation. Redox Biology, 65, 102810.
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3

Quantifying Oxidative Stress and Inflammation

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Serum levels of 8-OHdG were quantified with the 8-hydroxy 2 deoxyguanosine ELISA kit (Abcam, ab201734) according to the manufacturer’s instructions. IL-1β was measured in serum using the V-PLEX Mouse IL-1β Kit (Meso Scale Diagnostics, K152QPD-1), per the manufacturer’s instructions. The samples were read and analyzed by MSD QuickPlex SQ120 instrumentation and Workbench 4.0 Software (Meso Scale Diagnostics).
Marek-Iannucci S., Ozdemir A.B., Moreira D., Gomez A.C., Lane M., Porritt R.A., Lee Y., Shimada K., Abe M., Stotland A., Zemmour D., Parker S., Sanchez-Lopez E., Van Eyk J., Gottlieb R.A., Fishbein M.C., Karin M., Crother T.R., Rivas M.N, & Arditi M. (2021). Autophagy-mitophagy induction attenuates cardiovascular inflammation in a murine model of Kawasaki disease vasculitis. JCI Insight, 6(18), e151981.
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4

Plasma 8-Hydroxy-2'-Deoxyguanosine Measurement

Cited 3 times
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Plasma 8 OH-2dG measurement was performed by using the 8-hydroxy 2 deoxyguanosine ELISA Kit (Abcam, Cambridge, United Kingdom) (Surico et al., 2017 (link), 2019 (link); Grossini et al., 2018 (link), 2020 (link); De Cillà et al., 2019 (link), 2020 (link); Farruggio et al., 2019 (link), 2020 (link)), as described in Supplementary File 3. The 8 OH-2dG was detected using a wavelength of 450 nm.
Grossini E., Concina D., Rinaldi C., Russotto S., Garhwal D., Zeppegno P., Gramaglia C., Kul S, & Panella M. (2021). Association Between Plasma Redox State/Mitochondria Function and a Flu-Like Syndrome/COVID-19 in the Elderly Admitted to a Long-Term Care Unit. Frontiers in Physiology, 12, 707587.
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5

Quantifying Oxidative DNA Damage

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DNA was extracted using Isolate II Genomic DNA Kit (Bioline, London, UK), which was then denatured by heating at 95 °C for 5 min then rapidly cooled to 4 °C. Nuclease P1 enzyme (New England Biolabs, MA, USA) was added to the sample and incubated at 37 °C for 30 min then 75 °C for 10 min to break the DNA into single nucleotides, which were then dephosphorylated by incubation with FastAP Thermosensitive Alkaline Phosphatase (Invitrogen, MA, USA) at 37 °C for 10 min. The enzyme was deactivated by heating at 75 °C for 5 min. DNA oxidative damage was assessed by measuring the level of 8-hydroxy-2'-deoxyguanosine (8-OHdG) using 8-hydroxy 2 deoxyguanosine ELISA Kit (Abcam, Cambridge, UK) following the manufacturer’s instructions.
Nguyen L.T., Saad S., Shi Y., Wang R., Chou A.S., Gill A., Yao Y., Jarolimek W, & Pollock C.A. (2021). Lysyl oxidase inhibitors attenuate cyclosporin A-induced nephropathy in mouse. Scientific Reports, 11, 12437.
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6

Serum IgG and 8-OHDG Quantification

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Serum immunoglobulin G (IgG) levels were determined using the Rat IgG ELISA kit (Abcam, Cambridge, UK). The 8-hydroxy-2-deoxyguanosine (8-OHDG) level in rat serum and liver was determined using an 8-hydroxy 2 deoxyguanosine ELISA Kit (Abcam, Cambridge, UK), and the data were recorded on a Stat Fax 4300 chromate ELISA photometer (Awareness Technology, Palm City, FL, USA).
Kryl’skii E.D., Kravtsova S.E., Popova T.N., Matasova L.V., Shikhaliev K.S, & Medvedeva S.M. (2023). 6-Hydroxy-2,2,4-trimethyl-1,2-dihydroquinoline Demonstrates Anti-Inflammatory Properties and Reduces Oxidative Stress in Acetaminophen-Induced Liver Injury in Rats. Current Issues in Molecular Biology, 45(10), 8321-8336.
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7

Protein Extraction and Oxidative Stress Markers

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To extract the protein, SN was homogenized in Tris buffer (10 mM Tris-HCl, 1 mM EDTA, 0.32 M sucrose, and pH 7.8) as 10% (w/v) using a Teflon homogenizer (Glas-Col homogenizer system, Vernon Hills, USA), centrifuged at 20,000 × g for 10 min. The MDA, GSH, GSH-Px, SOD1, SOD2, and CAT contents in the SN were measured by using a commercially available kit (Jiancheng Bioengineering Institute, Nanjing, China) following the manufacturer's instructions. The 8-OHdG level was measured using 8-hydroxy 2-deoxyguanosine ELISA kit (Abcam, MA, USA), according to the manufacturer's protocol. The OD values were measured using a microplate reader and were expressed as percentage of the OD values of the control group.
Li R., Wang S., Li T., Wu L., Fang Y., Feng Y., Zhang L., Chen J, & Wang X. (2019). Salidroside Protects Dopaminergic Neurons by Preserving Complex I Activity via DJ-1/Nrf2-Mediated Antioxidant Pathway. Parkinson's Disease, 2019, 6073496.
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8

Quantifying Oxidative Stress via 8-OHdG ELISA

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The 8-hydroxy 2 deoxyguanosine ELISA Kit (Abcam, Cambridge, United Kingdom) was used to quantify the production of 8-OHdG as a marker of oxidative stress in the basolateral culture media 24 h post-exposure to aerosol. The assay was performed according to the manufacturer’s instructions. The optical density of each sample was measured with a microplate reader (Multiskan GO) at 450 nm. 8-OHdG production was quantified from a standard curve and expressed as a ratio to the negative control of air-exposed cells set to 1.
Mercier C., Pourchez J., Leclerc L, & Forest V. (2024). In vitro toxicological evaluation of aerosols generated by a 4th generation vaping device using nicotine salts in an air-liquid interface system. Respiratory Research, 25, 75.
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9

Biomarker Assessment in Oxidative Stress

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The S100B protein level was determined using an S100 Calcium Binding Protein B ELISA Kit (Cloud-Clone Corp., USA). The 8isoprostane concentration was assessed using a Rat 8-isoprostane ELISA Kit (Puda Scientific, China). The level of 8-hydroxy-2deoxyguanosine (8OHDG) was measured using an 8-hydroxy-2deoxyguanosine ELISA kit (Abcam, UK). The plates were washed with a Stat Fax 2600 washer (Awareness Technology, USA); the results were detected by a Stat Fax 4300 Chromate ELISA photometer (Awareness Technology, USA).
Kryl'skii E., Chupandina E., Popova T., Shikhaliev K., Mittova V., Popov S., Verevkin A., & Filin A. (2021). Neuroprotective effect of 6-hydroxy-2,2,4-trimethyl-1,2-dihydroquinoline mediated via regulation of antioxidant system and inhibition of inflammation and apoptosis in a rat model of cerebral ischemia/reperfusion. Biochimie, 186, 130-146.
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10

Quantitative ELISA for Plasma 8-OHdG

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The plasmatic 8-OHdG levels were measured using a competitive quantitative ELISA Kit (8-hydroxy-2-deoxyguanosine ELISA Kit, Abcam (Cambridge, UK)), according to the manufacturer’s instructions. The assay is based on the competition between 8-OHdG and an 8-OHdG acetylcholinesterase conjugate for a limited amount of 8-OHdG monoclonal antibody. The colorimetric intensity was determined spectrophotometrically in a SynergyTM multimode microplate reader, and its value was inversely proportional to the amount of free 8-OHdG in plasma.
Gonçalves A.C., Alves R., Baldeiras I., Marques B., Oliveiros B., Pereira A., Nascimento Costa J.M., Cortesão E., Mota Vieira L, & Sarmento Ribeiro A.B. (2021). DNA Methylation Is Correlated with Oxidative Stress in Myelodysplastic Syndrome—Relevance as Complementary Prognostic Biomarkers. Cancers, 13(13), 3138.
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