DNA Damage (8-OHdG) ELISA Kit

Mfr.No.	SKT-120
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, 8-OH-dG ELISA Kit ; 8OHG ELISA Kit ; 80G ELISA Kit ; 8 hydroxyguanine ELISA Kit ; 8-OHdG ELISA Kit ; DNA Damage ELISA Kit
Categories	ELISA Kits
Description	Colorimetric detection of 8-hydroxy-2-deoxy Guanosine
Field Of Use	Not for use in humans. Not for use in diagnostics or therapeutics. For in vitro research use only.
Gross Weight (kg)	2
Net Weight (g)	500
Reactivity	Species independent
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).
Specificity	Cross-reactivity: 8-Hydroxy-2-deoxy Guanosine (8-OHdG): 100%. 8-Hydroxy Guanosine (8-OHG): 23%. 8-Hydroxy Guanine (8-oxoG): 23%. Guanosine: <0.01%.
Size	1 Plate ; 1 vial/ 100uL ; 1 vial/75uL ; 1 vial/50mL ; 1 vial/13mL ; 1 vial/50mL ; 1 vial/13mL ; 1 vial/13mL ; 2 covers
Adr Code	Non-hazardous
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.
Assay Range	0.94 - 60 ng/mL
Assay Type	Competitive ELISA (Enzyme-linked Immunosorbent Assay)
Assay Type Abbreviation	Competitive
Component Number	SKC-120A ; SKC-120C ; SKC-120F ; SKC-0001 ; SKC-0002 ; SKC-0003 ; SKC-0004 ; SKC-0005 ; SKC-0009
Country Of Origin	Canada
Detection Method	Colorimetric Assay
Detection Range	0.94 - 60 ng/mL
Incubation Time	1 hour
Item	8-hydroxy-2-deoxy Guanosine : BSA Coated Plate ; 8-hydroxy-2-deoxy Guanosine Standard ; 8-hydroxy-2-deoxy Guanosine HRP Conjugated Monoclonal Antibody ; Sample and Standard Diluent ; 8-hydroxy-2-deoxy Guanosine Antibody Diluent ; Wash Buffer Concentrate ; TMB Substrate ; Stop Solution ; Plate Cover
Method Type	Urine, Cell Lysates, Plasma, Sample matrices
Number Of Samples	39 samples in duplicate
Platform	Microplate
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%.
Sensitivity	0.59 ng/mL
Species Reactivity Abbreviations	ALL
Species Reactivity Medium	Species Independent
Trademark	MOLECULAR SIGNATURE®

Research Area	Cancer Oxidative Stress Cell Signaling Post-translational Modifications Oxidation Cell Signaling Epigenetics and Nuclear Signaling DNA/RNA DNA Damage and Repair
Tarriff Code	3822.00.1090
Unspsc Code	12352203

Shipping Temperature

Blue Ice

Description

Trademark: MOLECULAR SIGNATURE®
Platform: Microplate
Utility: ELISA Kit for 8-OHdG detection in samples.
kit-overview:

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

Incubation Time: 1 hour
Shipping Temperature: Blue Ice
Gross Weight (kg): 2
Net Weight (g): 500
Stock Product: Y
Country of Origin: Canada

Storage

4ºC and -20ºC

Storage Temperature

4ºC and -20ºC

Stock Product

Utility

ELISA Kit for 8-OHdG detection in samples.

Reference	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.

Protocols Description	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.
Other Resources 1	Kit Booklet
Other Resources 2	MSDS
Other Resources 3	Calculations Worksheet