MOLECULAR SIGNATURE® DNA Damage (8-OHdG) ELISA kit

Colorimetric detection of 8-hydroxy-2-deoxy Guanosine

Catalog No. SKT-120

5.00 out of 5 based on 1 customer rating
Species Reactivity ALL
Sample Types Cell lysates, Plasma, Sample matrices, Urine
Rated 5.00 out of 5 based on 1 customer rating
(1 customer review)

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SKT-120_DNA_Damage_8-OHdG_ELISA_kit_Standard_Curve_Fig6.png
Chemical Equation of the Oxidation of Guanosine for the DNA Damage (8-OHdG) ELISA kit StressXpress - SKT-120Diagram of the 8-OHdG Competitive ELISA for the DNA Damage (8-OHdG) ELISA kit StressXpress - SKT-120Graph of the Urine Spike Assay for the DNA Damage (8-OHdG) ELISA kit StressXpress - SKT-120Diagram of the Preparation of the 8-OHdG Standards for the DNA Damage (8-OHdG) ELISA kit StressXpress - SKT-120Diagram of the Triplicate Sample Plate Format for the DNA Damage (8-OHdG) ELISA kit StressXpress - SKT-120Preview of the Calculations Worksheet for the DNA Damage (8-OHdG) ELISA kit StressXpress - SKT-120
Product Name DNA Damage (8-OHdG) ELISA kit
Description

Colorimetric detection of 8-hydroxy-2-deoxy Guanosine

Species Reactivity Species Independent
Platform Microplate
Sample Types Cell lysates, Plasma, Sample matrices, Urine
Detection Method Colorimetric Assay
Assay Type Competitive ELISA (Enzyme-linked Immunosorbent Assay)
Utility ELISA Kit for 8-OHdG detection in samples.
Sensitivity 0.59 ng/mL
Assay Range 0.94 - 60 ng/mL
Precision Intra-Assay Precision: Three samples of known concentration were assayed thirty times on one plate; the intra-assay coefficient of variation of the DNA Damage ELISA has been determined to be <5%. Inter-Assay Precision: Three samples of known concentration were assayed thirty times in three individual assays; the inter-assay coefficient of variation of the DNA Damage ELISA has been determined to be <5%.
Incubation Time 1 hour
Number of Samples 39 samples in duplicate
Other Resources Kit Booklet , MSDS , Calculations Worksheet

Properties

Storage Temperature 4ºC and -20ºC
Shipping Temperature Blue Ice
Product Type ELISA Kits
Assay Overview 1. Prepare standard and samples in the Sample and Standard Diluent. 2. Add 50 µL of prepared standards and samples in triplicate to appropriate wells. 3. Add 50 µL of the diluted antibody preparation to the appropriate wells. 4. Cover plate with Plate Cover and incubate at room temperature (20-25°C) for 1 hour. 5. Wash plate 4 times with 1X Wash Buffer. 6. Add 100 µL of TMB Substrate to each well. 7. Cover plate and develop the plate in the dark at room temperature for 30 minutes. 8. Add 100 µL of Stop Solution to each well. 9. Measure absorbance on a plate reader at 450 nm. 10. Plot the standard curve and calculate sample concentrations.
Kit Overview
Component No.ItemQuantity / Size
SKC-120A 8-hydroxy-2-deoxy Guanosine : BSA Coated Plate 1 Plate
SKC-120C 8-hydroxy-2-deoxy Guanosine Standard 1 vial/ 100uL
SKC-120F 8-hydroxy-2-deoxy Guanosine HRP Conjugated Monoclonal Antibody 1 vial/75uL
SKC-0001 Sample and Standard Diluent 1 vial/50mL
SKC-0002 8-hydroxy-2-deoxy Guanosine Antibody Diluent 1 vial/13mL
SKC-0003 Wash Buffer Concentrate 1 vial/50mL
SKC-0004 TMB Substrate 1 vial/13mL
SKC-0005 Stop Solution 1 vial/13mL
SKC-0009 Plate Cover  2 covers
Cite This Product DNA Damage (8-OHdG) ELISA Kit (StressMarq Biosciences Inc., Victoria BC CANADA, Catalog # SKT-120)

Biological Description

Alternative Names 8-OH-dG ELISA Kit, 8OHG ELISA Kit, 80G ELISA Kit, 8 hydroxyguanine ELISA Kit, 8-OHdG ELISA Kit, DNA Damage ELISA Kit
Research Areas Cancer, Cell Signaling, Oxidation, Oxidative Stress, Post-translational Modifications
Scientific Background 8-hydroxy-2-deoxy Guanosine (8-OH-dG) is produced by the oxidative damage of DNA by reactive oxygen and nitrogen species and serves as an established marker of oxidative stress (1-4). Hydroxylation of guanosine occurs in response to both normal metabolic processes and a variety of environmental factors (i.e., anything that increases reactive oxygen and nitrogen species). Increased levels of 8-OH-dG are associated with the aging process as well as with a number of pathological conditions including cancer, diabetes, and hypertension(5-9). In complex samples such as plasma, cell lysates, and tissues, 8-OH-dG can exist as either the free nucleoside or incorporated in DNA. Once the blood enters the kidney, free 8-OH-dG is readily filtered into the urine, while larger DNA fragments remain in the bloodstream. Because of the complexity of plasma samples, urine is a more suitable matrix for the measurement of free 8-OH-dG than plasma. Urinary levels of 8-OH-dG range between 2.7-13 ng/mg creatine, while plasma levels of free 8-OH-dG have been reported to be between 4-21 pg/ml as determined by LC-MS (10-11).
References 1. Maxey K.M., Maddipati K.R., Birkmeier J. (1992) J Clin Immunoassay 15: 116-120.
2. Pradelles P., Grassi J., Maclouf J. (1990) Methods Enzymol. 187: 24-34.
3. Maclouf J., Grassi J., Pradelles P. (1987) Dev Immunoassay Tech Meas eicosanoids.
4. Lin H., et al. (2004) Biochem J. 380: 541-548.
5. Bogdanov M.B., et al. (1999) Free Radic Biol Med. 27(5/6): 647-666.
6. Lee J., et al. (2005) Hypertension 45: 986-990.
7. Leinonen, J., et al. (1997) FEBSLett. 417: 150-152.
8. Endo K., et al. (2006) J. Atheroscler. Thromb. 13:68-75.
9. Kuo H., et al. (2007) Mutat Res. 631:62-68.
10. Shen J., et al. (2007) Cancer 109: 574-580.
11. Beckman K.B., Ames B.N. (1997) J Biol Chem 272: 19633-19636.
12. Epe B., et al. (1996) Nucleic Acids Res 24: 4105-4110.
13. Spencer J.P.E., et al. (1995) FEBS Lett 374: 233-236.
14. Floyd R.A. (1990) FASEB J 4: 2587-2597.

Product Images

<p>Typical Standard Curve for the DNA Damage (8-OHdG) ELISA kit (Enzyme-Linked Immunosorbent Assay) StressXpress® – SKT-120. Assay Type: Competitive ELISA. Detection Method: Colorimetric Assay. Assay Range: 0.94 – 60 ng/ml.</p>

Typical Standard Curve for the DNA Damage (8-OHdG) ELISA kit (Enzyme-Linked Immunosorbent Assay) StressXpress® – SKT-120. Assay Type: Competitive ELISA. Detection Method: Colorimetric Assay. Assay Range: 0.94 – 60 ng/ml.

<p>Chemical Equation of the Oxidation of Guanosine</p>

Chemical Equation of the Oxidation of Guanosine

<p>Diagram of the 8-OHdG Competitive ELISA</p>

Diagram of the 8-OHdG Competitive ELISA

<p>Urine Spike Assay</p>

Urine Spike Assay

<p>Diagram of the Preparation of the 8-OHdG Standards</p>

Diagram of the Preparation of the 8-OHdG Standards

<p>Diagram of the Triplicate Sample Plate Format</p>

Diagram of the Triplicate Sample Plate Format

<p>Preview of the Calculations Worksheet</p>

Preview of the Calculations Worksheet

Product Citations (43)

Other Citations

Association of Metabolic Syndrome and Oxidative DNA Damage in HIV/AIDS Patients.

Kolgiri, V., Nagar, V. and Patil, V. (2017) Ind J Clin Biochem. [Epub ahead of print].

PubMed ID: 28535982 Reactivity Human Applications: Plasma

Antiretroviral Therapy‑induced Insulin Resistance and Oxidative Deoxy Nucleic Acid Damage in Human Immunodeficiency Virus‑1 Patients.

VK Honnapurmath, V.W.P. (2017) Indian J Endocr Metab. 21:316-21.

PubMed ID: N/A Reactivity Human Applications: Blood

Urinary excretion of uric acid, allantoin, and 8-OH-Deoxyguanosine in uricase-knockout mice.

Inazawa, K. et al. (2016) Nucleosides Nucleotides Nucleic Acids. 35(10-12):559-565.

PubMed ID: 27906613 Reactivity Mouse Applications: Urine

Protein carbonyl content: a novel biomarker for aging in HIV/AIDS patients.

Kolgiri, V. and Patil, V.W. (2016) Braz J Infect Dis. pii: S1413-8670(16)30432-9.

PubMed ID: 27821249 Reactivity Human Applications: Plasma

Coagulin-l ameliorates TLR4 induced oxidative damage and immune response by regulating mitochondria and NOX-derived ROS.

Reddy, SS (2016) Toxicol Appl Pharmacol. 309:87-100.

PubMed ID: 27568862 Reactivity Mouse Applications: Liver homogenates

Preventive Effect of Resveratrol against Brain Mitochondria DNA Damage, Lipid Peroxidation, Inflammation and Seizures Induced by Kainic Acid in Mice.

Hussein, S. A., Abdel-mageid, A.D., Abd-Elhamed, O. M., Amin, A. and Al harthy, H.S. (2016) Int J Phar Sci. 6(4): 1634-1646

PubMed ID: N/A Reactivity Mouse Applications: Brain tissue

Neuroprotective Effect of Curcumin on Kainic Acid Model of Epilepsy in Male Swiss Albino Mice.

Hussein, S. A., Abdel-mageid, A.D., Abd-Elhamed, O. M., Amin, A. and Al harthy, H.S. (2016) Int J Chem Nat Sci. 4(4): 447-460.

PubMed ID: N/A Reactivity Mouse Applications: Brain tissue

DNA damage and the activation of the p53 pathway mediate alterations in metabolic and secretory functions of adipocytes.

Vergoni, B. et al. (2016) Diabetes.

PubMed ID: 27388216 Reactivity Human Applications: Adipocytes

Podocyte-specific Nox4 deletion affords renoprotection in a mouse model of diabetic nephropathy.

Jha, J.C. et al. (2015) Diabetologia. 59(2):379-89.

PubMed ID: 26508318 Reactivity Mouse Applications: Urine

Does chronic raise of metal ion levels induce oxidative DNA damage and hypoxia-like response in patients with metal-on-metal hip resurfacing?

Savarino, L. et al. (2015) J Biomed Mater Res B Appl Biomater. 105(2):460-466.

PubMed ID: 26477446 Reactivity Human Applications: Serum

Higher urinary Levels of 8-hydroxy-2'-deoxyguanosine are associated with a worse RANKL/OPG ratio in postmenopausal women with osteopenia.

Cervellati, C. et al. (2015) Oxid Med Cell Long. 2016:6038798.

PubMed ID: 26635910 Reactivity Human Applications: Urine

Turmerone enriched standardized Curcuma longa extract alleviates LPS induced inflammation and cytokine production by regulating TLR4–IRAK1–ROS–MAPK–NFκB axis.

Rana, M. et al. (2015) J Functional Foods. 16 (2015): 152–163.

PubMed ID: N/A Reactivity Mouse Applications: Liver homogenates

Synthesis, characterization and biological activity of some unsymmetrical Schiff base transition metal complexes.

Esmadi, F.T. et al. (2015) Drug Chem Toxicol. :1-7.

PubMed ID: 25791998 Reactivity Human Applications: Lymphocytes

Late-intervention study with ebselen in an experimental model of type 1 diabetic nephropathy.

Tan, S.M., Sharma, A., Stefanovic, N. and de Haan, J.B. (2015) Free Radic Res. 49(3):219-27.

PubMed ID: 25465090 Reactivity Mouse Applications: Urine

Immediate and delayed effects of growth conditions on ageing parameters in nestling zebra finches.

Reichert, S. et al. (2015) J Exp Biol. 218(Pt 3):491-9.

PubMed ID: 25524985 Reactivity Zebra finch Applications: Blood cells

Elevation impacts the balance between growth and oxidative stress in coal tits.

Stier, A., et al. (2014) Oecologia. 175(3):791-800.

PubMed ID: 24805201 Reactivity Bird (coal tits) Applications: Red blood cells

Evaluation of vitamin B12 effects on DNA damage induced by paclitaxel.

Alzoubi, K., Khabour O., Khader M., Mhaidat N. and Al-Azzam, S. (2014) Drug Chem Toxicol. 37(3):276-80.

PubMed ID: 24215581 Reactivity Human Applications: Blood

Disruption of pro-oxidant and antioxidant systems with elevated expression of the ubiquitin proteosome system in the cachectic heart muscle of nude mice.

Hinch, E. C. A., Sullivan-Gunn, M. J., Vaughan V. C., McGlynn M. A., Lewandowski, Paul A. (2013) J Cachexia Sarcopenia Muscle. 4(4):287-93.

PubMed ID: 24030522 Reactivity Mouse Applications: Tumor tissue

Oxidative Damage of DNA Confers Resistance to Cytosolic Nuclease TREX1 Degradation and Potentiates STING-Dependent Immune Sensing.

Gehrke, N. et al. (2013) Immunity. 39(3):482-95.

PubMed ID: 23993650 Reactivity Mouse Applications: RMA cells (T-cell tumor cells)

Differential effects of docosahexanoic acid (DHA) on preterm and term placental pro-oxidant/anti-oxidant balance.

Stark, M. et al. (2013) Reproduction. 146(3):243-51.

PubMed ID: 23813449 Reactivity Human Applications: Placenta

The Modified Selenenyl Amide, M-hydroxy Ebselen, Attenuates Diabetic Nephropathy and Diabetes-Associated Atherosclerosis in ApoE/GPx1 Double Knockout Mice.

Tan, S.M., et al. (2013) PLoS ONE. 8(7): e69193.

PubMed ID: 23874911 Reactivity Mouse Applications: Urine (diabetic and non-diabetic)

TiO2 Nanoparticle Exposure and Illumination during Zebrafish Development: Mortality at Parts per Billion Concentrations.

Bar-Ilan, O. et al. (2013) Environ Sci Technol. 47 (9): 4726-4733.

PubMed ID: 23510150 Reactivity Zebrafish Applications: DNA extract

Poor sleep in PCOS; is melatonin the culprit?

Shreeve, N. et al. (2013) Hum Reprod. 28 (5): 1348-1353.

PubMed ID: 23438443 Reactivity Human Applications: Urine

Grasshoppers' adaptation to elevated radioactivity in the Chernobyl exclusion zone.

Mortensen, L.H. (2013) Roskilde University Dissertation

PubMed ID: N/A Reactivity Grasshopper Applications: Intestines

Consumption of a low glycaemic index diet in late life extends lifespan of Balb/c mice with differential effects on DNA damage.

Nankervis, S.A., Mitchell, J.M., Charchar, F.J., McGlynn, M.A. and Lewandowski, P.A. (2013) Longev Healthspan. 2(1):4.

PubMed ID: 24472560 Reactivity Mouse Applications: WBC's

Light-induced cell detachment for cell sheet technology.

Hong, Y. et al. (2013) Biomaterials. 34 (1): 11-18.

PubMed ID: 23069710 Reactivity Mouse Applications: MC3T3-E1 cells

Puerarin prevents high glucose-induced apoptosis of Schwann cells by inhibiting oxidative stress.

Wu, Y., Xue, B., Li, X. and Liu, H. (2012) Neural Regen Res. 7 (33): 2583-2591.

PubMed ID: 25368634 Reactivity Mouse Applications: Schwann cells

The protective effect of Alpha lipoic acid on Schwann cells exposed to constant or intermittent high glucose.

Sun, L. et al. (2012) Biochem Pharmacol. 84 (7): 961-973.

PubMed ID: 22796564 Reactivity Mouse Applications: Schwann cells

Ginsenoside Rb1 relieves glucose fluctuation induced oxidative stress and apoptosis in Schwann cells.

Xue, B. et al. (2012) Neural Regen Res. 2012 (30): 2340-2346.

PubMed ID: 25538758 Reactivity Mouse Applications: Schwann cells

Eicosapentaenoic Acid and Oxypurinol in the Treatment of Muscle Wasting in a Mouse Model of Cancer Cachexia.

Vaughan, V.C., Sullivan-Gunn, M., Hinch, E., Martin, P., Lewandowski, P.A. (2012) PLoS ONE. 7(9): e45900.

PubMed ID: 23029301 Reactivity Mouse Applications: Gastrocnemius muscle

Titanium dioxide nanoparticles produce phototoxicity in the developing zebrafish.

Bar-Ilan, O. et al. (2012) Nanotoxicology. 6 (6): 670-679.

PubMed ID: 21830861 Reactivity Zebrafish Applications: Embryos

Evaluation of vitamin B12 effects on DNA damage induced by pioglitazone.

Alzoubi, K., Khabour, O., Hussain, N., Al-azzam, S. and Mhaidat, N. (2012) Mutat Res. 748 (1-2): 48-51.

PubMed ID: 22790087 Reactivity Human Applications: Blood

Protective Effects of Salvianolic Acid B on Schwann Cells Apoptosis Induced by High Glucose.

Sun, L. et al. (2012) Neurochemical Res. 37 (5): 996-1010.

PubMed ID: 22252725 Reactivity Mouse Applications: Schwann cells

Oxidative Stress in HPV-Driven Viral Carcinogenesis: Redox Proteomics Analysis of HPV-16 Dysplastic and Neoplastic Tissues.

De Marco, F. et al. (2012) PLoS ONE. 7(3): e34366.

PubMed ID: 22470562 Reactivity Human Applications: Ecto-cervical tissue

Inhibitory effects of Salvianolic acid B on apoptosis of Schwann cells and its mechanism induced by intermittent high glucose.

Sun, L. et al. (2012) Life Sci. 90 (3-4): 99-108.

PubMed ID: 22036624 Reactivity Mouse Applications: Schwann cells

A qPCR-based assay to quantify oxidized guanine and other FPG-sensitive base lesions within telomeric DNA.

O'Callaghan, N., Baack, N., Sharif, R. and French, M. (2011) Biotechniques. 51 (6): 403-411.

PubMed ID: 22150331 Reactivity Human Applications: Spleen B lymphoblastoid cells

The effect of cocoa supplementation on hepatic steatosis, reactive oxygen species and LFABP in a rat model of NASH.

Janevski, M., Antonas, K.N., Sullivan-Gunn, M.J., McGlynn, M.A. and Lewandowski, P.A. (2011) Comp Hepatol. 10(1):10.

PubMed ID: 22081873 Reactivity Rat Applications: Liver tissue

Reduction of Oxidative Damage Reflects a Better Kidney Transplantation Outcome. 

La Manna, G. et al.  (2011) Am J Nephrol. 34, 496-504. 

PubMed ID: 22041478 Reactivity Human Applications: PBMC

Integrated exposure assessment of sewage workers to genotoxicants: an urinary biomarker approach and oxidative stress evaluation.

Zabadi, H.A. et al. (2011) Environ Health. 10 (23).

PubMed ID: 21435260 Reactivity Human Applications: Urine

Transgenic Mice Expressing Cyclooxygenase-2 in Hepatocytes Reveal a Minor Contribution of This Enzyme to Chemical Hepatocarcinogenesis.

Izquierdo, C.L. et al. (2011) Am J Pathol. 178 (3): 1361-1373.

PubMed ID: 21356386 Reactivity Mouse Applications: COX-2 Tg cells

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