Colorimetric detection of 8-hydroxy-2-deoxy Guanosine__Colorimetric detection of 8-hydroxy-2-deoxy Guanosine Tipifarnib
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
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)
References PubMed ID：:http://www.ncbi.nlm.nih.gov/pubmed/19120987
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.
Colorimetric detection of 8-hydroxy-2-deoxy Guanosine
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%.
Kit Booklet
,
MSDS
,
Calculations Worksheet
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.
| 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 |
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.