DNA Damage (8-OHdG) ELISA kit

Colorimetric detection of 8-hydroxy-2-deoxy Guanosine

Catalog No. SKT-120

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Species Reactivity , ALL
Sample Types Cell lysates, Plasma, Sample matrices, Urine

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SKU: SKT-120 Categories: , .

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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-120Preview of the Calculations Worksheet for the DNA Damage (8-OHdG) ELISA kit StressXpress - SKT-120

Overview

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 Components
Component No. Item Quantity / 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: 16-120.
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

Chemical Equation of the Oxidation of Guanosine

Urine Spike Assay

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.

Diagram of the Preparation of the 8-OHdG Standards

Diagram of the 8-OHdG Competitive ELISA

Preview of the Calculations Worksheet

Product Citations

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