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Stock availability
In Stock
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Kit s description
Colorimetric detection of 8-hydroxy-2-deoxy Guanosine
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Protein target
DNA Damage (8-OHdG)
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Scientific context
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).
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Primary research area
Cancer, Oxidative Stress, Cell Signaling, Post-Translational Modifications, Oxidation
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Category
Assay ELISA Kits
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Other name
8-OH-dG ELISA Kit, 8OHG ELISA Kit, 80G ELISA Kit, 8 hydroxyguanine ELISA Kit, 8-OHdG ELISA Kit, DNA Damage ELISA Kit
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Brand name
MOLECULAR SIGNATURE®
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Detection system
Colorimetric Assay
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Assay format
Competitive ELISA (Enzyme-linked Immunosorbent Assay)
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Main intended usage
ELISA Kit for 8-OHdG detection in samples.
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Brief protocol
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.
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Kit contents
96 well microplate for ELISA detection of 8-hydroxy-2-deoxy Guanosine (8-OH-dG)
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Assay 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
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Assay cross reactivity
Species Independent
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Samples to be used with this kit
Urine, Cell Lysates, Plasma, Sample matrices
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Maximum samples to be used with this kit
39 samples in duplicate
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Assay duration
1 hour
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Kit s sensitivity
0,59 ng/mL
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Assay detection limit
0,94 - 60 ng/mL
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Storage recommendations
4°C and -20°C
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Shipping recommendations
Blue Ice
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Bibliography
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.
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Release date
1-Aug-2015
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PubMed number
27906613, 27821249, 27568862, 27388216, 27084692, 26508318, 26477446, 26635910, 25791998, 25465090, 25524985, 24740568, 24805201, 24215581, 24343823, 24030522, 23993650, 23813449, 23874911, 23510150, 23438443, 24472560, 23069710, 25368634, 22796564, 25538758, 23029301, 21830861, 22790087, 22252725, 22470562, 22036624, 22150331, 22081873, 22041478, 21435260, 21356386
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Tested applications
EIA, ELISA
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Tested reactivity
Mouse, Human, Mouse, Human, Human, Mouse, Human, Human, Human, Mouse, Zebra finch, Mice, Bird (coal tits), Human, Human, Mouse, Mouse, Human, Mouse, Zebrafish, Human, Mouse, Mouse, Mouse, Mouse, Mouse, Mouse, Zebrafish, Human, Mouse, Human, Mouse, Human, Rat, Human, Human, Mouse
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Resources available upon request
Kit Booklet, MSDS, Calculations Worksheet
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Product image link
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Product image legend
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. | Chemical Equation of the Oxidation of Guanosine | Diagram of the 8-OHdG Competitive ELISA | Urine Spike Assay | Diagram of the Preparation of the 8-OHdG Standards | Diagram of the Triplicate Sample Plate Format | Preview of the Calculations Worksheet Graph of the Typical Standard Curve for the DNA Damage (8-OHdG) ELISA kit StressXpress - SKT-120 | Chemical Equation of the Oxidation of Guanosine for the DNA Damage (8-OHdG) ELISA kit StressXpress - SKT-120 | Diagram of the 8-OHdG Competitive ELISA for the DNA Damage (8-OHdG) ELISA kit StressXpress - SKT-120 | Graph of the Urine Spike Assay for the DNA Damage (8-OHdG) ELISA kit StressXpress - SKT-120 | Diagram of the Preparation of the 8-OHdG Standards for the DNA Damage (8-OHdG) ELISA kit StressXpress - SKT-120 | Diagram of the Triplicate Sample Plate Format for the DNA Damage (8-OHdG) ELISA kit StressXpress - SKT-120 | Preview of the Calculations Worksheet for the DNA Damage (8-OHdG) ELISA kit StressXpress - SKT-120
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Warnings
Non-hazardous
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Country of production
Canada
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Total weight kg
2
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Net weight g
500
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Properties
E05 478 566 350 170 or Enzyme-Linked Immunosorbent Assays, E05 478 566 350 170 or Enzyme-Linked Immunosorbent Assays
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Test
ELISA Enzyme-linked immunosorbent assays Code 90320007 SNOMED