Typical Testing Data/Standard Curve (for reference only)
OxiSelect Protein Carbonyl ELISA Kit
OxiSelect Protein Carbonyl ELISA Kit (1 plate)
Synonyms
OxiSelect Protein Carbonyl; OxiSelect Protein Carbonyl ELISA Kit (1 plate); OxiSelect Protein Carbonyl elisa kit
Preparation and Storage
Upon receipt, aliquot and store both the Reduced and Oxidized BSA Standards at -20 degree C to avoid multiple freeze/thaw cycles. Store all other components at 4 degree C until their expiration dates.
Typical Testing Data/Standard Curve (for reference only)
Typical Testing Data/Standard Curve (for reference only)
Related Product Information for OxiSelect Protein Carbonyl elisa kit
Principle of the Assay: BSA standards or protein samples (10 ug/mL) are adsorbed onto a 96-well plate for 2 hrs at 37 degree C. The protein carbonyls present in the sample or standard are derivatized to DNP hydrazone and probed with an anti-DNP antibody, followed by an HRP conjugated secondary antibody. The protein carbonyl content in unknown sample is determined by comparing with a standard curve that is prepared from predetermined reduced and oxidized BSA standards.
Background/Introduction: Protein oxidation is defined as the covalent modification of a protein induced either directly by reactive oxygen species (ROS) or indirectly by reaction with secondary by-products of oxidative stress. Oxidative modification of proteins can be induced in vitro by a wide array of pro-oxidant agents and occurs in vivo during aging and in certain disease conditions. There are numerous types of protein oxidative modification. The most common products of protein oxidation in biological samples are the protein carbonyl derivatives of Pro, Arg, Lys, and Thr. These derivatives are chemically stable and serve as markers of oxidative stress for most types of ROS. Many of the current assays involve derivatization of the carbonyl group with dinitrophenylhydrazine (DNPH), followed by immunobloting with an anti-DNP antibody. The Protein Carbonyl ELISA was first developed by Buss and co-workers, the protein samples (>4 mg/mL) react with DNPH and then adsorb to wells of an ELISA plate before probe with anti-DNPH antibody. In their method, protein samples containing low amounts of protein must be concentrated to at least 4 mg/mL by TCA precipitation. However, TCA precipitation results a 20% loss of the total carbonyl values, and loss of protein during precipitation is also expected. In OxiSelect™ Protein Carbonyl ELISA Kit, protein samples are first allowed to adsorb to wells of a 96-well plate and then react with DNPH. There is no need to concentrate protein in experimental and clinical samples with low amounts of protein (< 4 mg/mL) and the kit requires protein sample as little as 10 ug/mL. The OxiSelect™ Protein Carbonyl ELISA Kit is an enzyme immunoassay developed for rapid detection and quantitation of protein carbonyls. The quantity of protein carbonyls in protein sample is determined by comparing its absorbance with that of a known reduced/oxidized BSA standard curve. Each kit provides sufficient reagents to perform up to 96 assays, including standard curve and unknown protein samples.
Background/Introduction: Protein oxidation is defined as the covalent modification of a protein induced either directly by reactive oxygen species (ROS) or indirectly by reaction with secondary by-products of oxidative stress. Oxidative modification of proteins can be induced in vitro by a wide array of pro-oxidant agents and occurs in vivo during aging and in certain disease conditions. There are numerous types of protein oxidative modification. The most common products of protein oxidation in biological samples are the protein carbonyl derivatives of Pro, Arg, Lys, and Thr. These derivatives are chemically stable and serve as markers of oxidative stress for most types of ROS. Many of the current assays involve derivatization of the carbonyl group with dinitrophenylhydrazine (DNPH), followed by immunobloting with an anti-DNP antibody. The Protein Carbonyl ELISA was first developed by Buss and co-workers, the protein samples (>4 mg/mL) react with DNPH and then adsorb to wells of an ELISA plate before probe with anti-DNPH antibody. In their method, protein samples containing low amounts of protein must be concentrated to at least 4 mg/mL by TCA precipitation. However, TCA precipitation results a 20% loss of the total carbonyl values, and loss of protein during precipitation is also expected. In OxiSelect™ Protein Carbonyl ELISA Kit, protein samples are first allowed to adsorb to wells of a 96-well plate and then react with DNPH. There is no need to concentrate protein in experimental and clinical samples with low amounts of protein (< 4 mg/mL) and the kit requires protein sample as little as 10 ug/mL. The OxiSelect™ Protein Carbonyl ELISA Kit is an enzyme immunoassay developed for rapid detection and quantitation of protein carbonyls. The quantity of protein carbonyls in protein sample is determined by comparing its absorbance with that of a known reduced/oxidized BSA standard curve. Each kit provides sufficient reagents to perform up to 96 assays, including standard curve and unknown protein samples.
Product Categories/Family for OxiSelect Protein Carbonyl elisa kit
References
1. Cadenas, E., Boveris, A., Ragan, CI., and Stoppani, AO. (1977). Archives of Biochemistry & Biophysics 180:248257.
2. Wakeyama, H., Takeshige, K., Takayanagi, R., and Minakami, S. (1982). Biochem J. 205:593601.
3. Talent, JM., Kong, Y., and Gracy, RW. (1998). Anal. Biochem. 263:3138.
4. Buss, H, Chan, TP, Sluis, KB, Domigan, NM, and Winterbourn, CC. (1985) Free Radic Biol Med. 23:361-6.
5. Reznick, AZ., and Packer, L. (1994) Methods Enzymol. 233:263-357.
1. Westenbrink, B. D. et al. (2015). Mitochondrial reprogramming induced by CaMKIIdelta mediates hypertrophy decompensation. Circ Res. 116:e28-e39.
2. Alway, S. E. et al. (2015). Green tea extract attenuates muscle loss and improves muscle function during disuse, but fails to improve muscle recovery following unloading in aged rats. J. Appl Physiol (1985). 118:319-330.
3. Yu, J. et al. (2015). Influence of chlorpyrifos oxon on the development and progression of alzheimer's disease in amyloid precursor protein transgenic mice. Neuroimmunology and Neuroinflammation 2:31.
4. Shen, Y. B. et al. (2014). Effect of feed grade L-methionine on growth performance and gut health in nursery pigs compared with conventional DL-methionine. J Anim Sci. 92:5530-5539.
5. Sosnowska, D. et al. (2014). A heart that beats for 500 years: age-related changes in cardiac proteasome activity, oxidative protein damage and expression of heat shock proteins, inflammatory factors, and mitochondrial complexes in arctica islandica, the longest-living noncolonial animal. J Gerontol A Biol Sci Med Sci. 69:1448-1461.
6. Al-Khalili, L. et al. (2014). Proteasome inhibition in skeletal muscle cells unmasks metabolic derangements in type 2 diabetes. Am J Physiol Cell Physiol. 307:C774-C787.
7. Ravikumar, P. et al. (2014). alpha-Klotho protects against oxidative damage in pulmonary epithelia. Am J Physiol Lung Cell Mol Physiol. 307:L566-575.
8. Liu, C. et al. (2014). Decreased DJ-1 leads to impaired Nrf2-regulated antioxidant defense and increased UV-A-induced apoptosis in corneal endothelial cells. Invest Ophthalmol Vis Sci. 55:5551-5560.
9. Farmer, C. et al. (2014). Effects of the plant extract silymarin on prolactin concentrations, mammary gland development, and oxidative stress in gestating gilts. J Anim Sci. 92:2922-2930.
10. DuPont, J. J. et al. (2014). NADPH Oxidase-derived reactive oxygen species contribute to impaired cutaneous microvascular function in chronic kidney disease. Am J Physiol Renal Physiol. 306:F1499-F1506.
11. Olesen, J. et al. (2014). Exercise training, but not resveratrol, improves metabolic and inflammatory status in skeletal muscle of aged men. J. Physiol. 592:1873-1886.
12. Vulesevic, B. et al. (2014). Glyoxalase-1 overexpression in bone marrow cells reverses defective neovascularization in STZ-induced diabetic mice. Cardiovasc Res. 101:306-316.
13. Zhao, Y. et al. (2013). Effect of social ranks and gestation housing systems on oxidative stress status, reproductive performance, and immune status of sows. J Anim Sci. 91:5848-5858.
14. Sosnowska, D. et al. (2013). A heart that beats for 500 years: age-related changes in cardiac proteasome activity, oxidative protein damage and expression of heat shock proteins, inflammatory factors, and mitochondrial complexes in arctica islandica, the longest-living noncolonial animal. J Gerontol A Biol Sci Med Sci. 10.1093/gerona/glt201.
15. Di Cara, F. et al. (2013). Invadolysin, a conserved lipid-droplet-associated metalloproteinase, is required for mitochondrial function in drosophila. J. Cell Sci. 126:4769-4781.
16. Dai, D. F. et al. (2013).Global proteomics and pathway analysis of pressure-overload-induced heart failure and its attenuation by mitochondrial-targeted peptides. Circ Heart Fail. 6:1067-1076.
17. Ungvari, Z. et al. (2013). Testing predictions of the oxidative stress hypothesis of aging using a novel invertebrate model of longevity: the giant clam (Tridacna derasa). J Gerontol A Biol Sci Med Sci. 68:359-367.
18. Young, K. et al. (2013). Each to their own: skeletal muscles of different function use different biochemical strategies during aestivation at high temperature. J.Exp. Biol. 216:1012-1024.
19. Fishman, J. et al. (2012). Oxidative modification of the intestinal mucus layer is a critical but unrecognized component of trauma hemorrhagic shock-induced gut barrier failure. Am J Physiol Gastrointest Liver Physiol. 304:G57-G63.
20. Murakami.Y. et al. (2012). Receptor interacting protein kinase mediates necrotic cone but not rod cell death in a mouse model of inherited degeneration. PNAS. 10.1073/pnas.1206937109.
21. Kang, K.A. et al. (2012). Baicalein inhibits oxidative stress-induced cellular damage via antioxidant effects. Toxicology and Industrial Health. 28:412-421.
22. Gannon, A.M. et al. (2012). Cigarette smoke exposure leads to follicle loss via an alternative ovarian cell death pathway in a mouse model. Toxicol. Sci. 125:274-284.
23. Tighe, R.M. et al. (2011). Ozone inhalation promotes CX3CR1-dependent maturation of resident lung macrophages that limit oxidative stress and inflammation. J. Immunol. 187:4800-4808.
24. Tang, H. et al. (2011). Intrinsic apoptosis in mechanically ventilated human diaphragm: linkage to a novel Fos/FoxO1/Stat3-Bim axis. FASEB J. 25:2921-2936.
25. Roh, M.I. et al. (2011). Edaravone, an ROS scavenger, ameliorates photoreceptor cell death after experimental retinal detachment. Invest. Ophthalmol. Vis. Sci. 52:3825-3831.
26. Robinson, C.K. et al. (2011). A major role for nonenzymatic antioxidant processes in the radioresistance of Halobacterium salinarum. J. Bacteriol. 193:1653-1662.
27. Dai, D.F. et al. (2011). Mitochondrial targeted antioxidant peptide ameliorates hypertensive cardiomyopathy. J. Am. Coll. Cardiol. 58:73-82.
28. Thompson, C.M. et al. (2011). Investigation of the mode of action underlying the tumorigenic response induced in B6C3F1 mice exposed orally to hexavalent chromium. Toxicol. Sci. 123:58-70.
29. Ungvari, Z. et al. (2011). Extreme longevity is associated with increased resistance to oxidative stress in Arctica islandica, the longest-living non-colonial animal. J. Gerontol A Biol Sci Med Sci. 10.1093/gerona/glr044
30. Labbé, A. et al. (2010). Resveratrol improves insulin resistance hyperglycemia and hepatosteatosis but not hypertriglyceridemia, inflammation, and life span in a mouse model for Werner Syndrome. J. Gerontol. A Biol Sci Med. Sci. 10.1093/gerona/glq184.
31. Nolin, T.D. et al (2010). Multiple-dose pharmacokinetics and pharmacodynamics of N-acetylcysteine in patients with end-stage renal disease. Clin. J. Am. Soc. Nephrol. 5:1588-1594.
32. Kang, K.A. et al. (2010). KIOM-4 protects against oxidative stress-induced mitochondrial damage in pancreatic beta-Cells via its antioxidant effects. Evid. Based Complement. Altern. Med. 10.1093/ecam/neq007.
33. Cho, H-Y. et al. (2009). Antiviral activity of Nrf2 in a murine model of respiratory syncytial virus (RSV) disease. Am. J. Respir. Crit. Care Med. 179:138-150.
34. Neretti, N. et al. (2009). Long lived Indy induces reduced mitochondrial reactive oxygen species production and oxidative damage. PNAS 106:2277-2282.
35. Maity, P. et al. (2008). Indomethacin, a non-steroidal anti-inflammatory drug, develops gastropathy by inducing reactive oxygen species-mediated mitochondrial pathology and associated apoptosis in gastric mucosa: a novel role of mitochondrial aconitase oxidation. J. Biol. Chem. 284:3058-3068.
36. Kim, S-H. et al. (2008). Protective effect of esculetin against oxidative stress-induced cell damage via scavenging reactive oxygen species. Acta Pharmacol. Sinica 29(11):1319-1326.
37. Paromov, V. et al. (2008). The influence of N-acetyl-L-cysteine on oxidative stress and nitric oxide synthesis in stimulated macrophages treated with a mustard gas analogue. BMC Cell Biology 9:33.
38. Lowry, M. et al. (2007). Lung lining fluid glutathione attenuates IL-13 induced asthma. Am. J. Respir. Cell Mol. Biol. 38(5):509-516.
2. Wakeyama, H., Takeshige, K., Takayanagi, R., and Minakami, S. (1982). Biochem J. 205:593601.
3. Talent, JM., Kong, Y., and Gracy, RW. (1998). Anal. Biochem. 263:3138.
4. Buss, H, Chan, TP, Sluis, KB, Domigan, NM, and Winterbourn, CC. (1985) Free Radic Biol Med. 23:361-6.
5. Reznick, AZ., and Packer, L. (1994) Methods Enzymol. 233:263-357.
1. Westenbrink, B. D. et al. (2015). Mitochondrial reprogramming induced by CaMKIIdelta mediates hypertrophy decompensation. Circ Res. 116:e28-e39.
2. Alway, S. E. et al. (2015). Green tea extract attenuates muscle loss and improves muscle function during disuse, but fails to improve muscle recovery following unloading in aged rats. J. Appl Physiol (1985). 118:319-330.
3. Yu, J. et al. (2015). Influence of chlorpyrifos oxon on the development and progression of alzheimer's disease in amyloid precursor protein transgenic mice. Neuroimmunology and Neuroinflammation 2:31.
4. Shen, Y. B. et al. (2014). Effect of feed grade L-methionine on growth performance and gut health in nursery pigs compared with conventional DL-methionine. J Anim Sci. 92:5530-5539.
5. Sosnowska, D. et al. (2014). A heart that beats for 500 years: age-related changes in cardiac proteasome activity, oxidative protein damage and expression of heat shock proteins, inflammatory factors, and mitochondrial complexes in arctica islandica, the longest-living noncolonial animal. J Gerontol A Biol Sci Med Sci. 69:1448-1461.
6. Al-Khalili, L. et al. (2014). Proteasome inhibition in skeletal muscle cells unmasks metabolic derangements in type 2 diabetes. Am J Physiol Cell Physiol. 307:C774-C787.
7. Ravikumar, P. et al. (2014). alpha-Klotho protects against oxidative damage in pulmonary epithelia. Am J Physiol Lung Cell Mol Physiol. 307:L566-575.
8. Liu, C. et al. (2014). Decreased DJ-1 leads to impaired Nrf2-regulated antioxidant defense and increased UV-A-induced apoptosis in corneal endothelial cells. Invest Ophthalmol Vis Sci. 55:5551-5560.
9. Farmer, C. et al. (2014). Effects of the plant extract silymarin on prolactin concentrations, mammary gland development, and oxidative stress in gestating gilts. J Anim Sci. 92:2922-2930.
10. DuPont, J. J. et al. (2014). NADPH Oxidase-derived reactive oxygen species contribute to impaired cutaneous microvascular function in chronic kidney disease. Am J Physiol Renal Physiol. 306:F1499-F1506.
11. Olesen, J. et al. (2014). Exercise training, but not resveratrol, improves metabolic and inflammatory status in skeletal muscle of aged men. J. Physiol. 592:1873-1886.
12. Vulesevic, B. et al. (2014). Glyoxalase-1 overexpression in bone marrow cells reverses defective neovascularization in STZ-induced diabetic mice. Cardiovasc Res. 101:306-316.
13. Zhao, Y. et al. (2013). Effect of social ranks and gestation housing systems on oxidative stress status, reproductive performance, and immune status of sows. J Anim Sci. 91:5848-5858.
14. Sosnowska, D. et al. (2013). A heart that beats for 500 years: age-related changes in cardiac proteasome activity, oxidative protein damage and expression of heat shock proteins, inflammatory factors, and mitochondrial complexes in arctica islandica, the longest-living noncolonial animal. J Gerontol A Biol Sci Med Sci. 10.1093/gerona/glt201.
15. Di Cara, F. et al. (2013). Invadolysin, a conserved lipid-droplet-associated metalloproteinase, is required for mitochondrial function in drosophila. J. Cell Sci. 126:4769-4781.
16. Dai, D. F. et al. (2013).Global proteomics and pathway analysis of pressure-overload-induced heart failure and its attenuation by mitochondrial-targeted peptides. Circ Heart Fail. 6:1067-1076.
17. Ungvari, Z. et al. (2013). Testing predictions of the oxidative stress hypothesis of aging using a novel invertebrate model of longevity: the giant clam (Tridacna derasa). J Gerontol A Biol Sci Med Sci. 68:359-367.
18. Young, K. et al. (2013). Each to their own: skeletal muscles of different function use different biochemical strategies during aestivation at high temperature. J.Exp. Biol. 216:1012-1024.
19. Fishman, J. et al. (2012). Oxidative modification of the intestinal mucus layer is a critical but unrecognized component of trauma hemorrhagic shock-induced gut barrier failure. Am J Physiol Gastrointest Liver Physiol. 304:G57-G63.
20. Murakami.Y. et al. (2012). Receptor interacting protein kinase mediates necrotic cone but not rod cell death in a mouse model of inherited degeneration. PNAS. 10.1073/pnas.1206937109.
21. Kang, K.A. et al. (2012). Baicalein inhibits oxidative stress-induced cellular damage via antioxidant effects. Toxicology and Industrial Health. 28:412-421.
22. Gannon, A.M. et al. (2012). Cigarette smoke exposure leads to follicle loss via an alternative ovarian cell death pathway in a mouse model. Toxicol. Sci. 125:274-284.
23. Tighe, R.M. et al. (2011). Ozone inhalation promotes CX3CR1-dependent maturation of resident lung macrophages that limit oxidative stress and inflammation. J. Immunol. 187:4800-4808.
24. Tang, H. et al. (2011). Intrinsic apoptosis in mechanically ventilated human diaphragm: linkage to a novel Fos/FoxO1/Stat3-Bim axis. FASEB J. 25:2921-2936.
25. Roh, M.I. et al. (2011). Edaravone, an ROS scavenger, ameliorates photoreceptor cell death after experimental retinal detachment. Invest. Ophthalmol. Vis. Sci. 52:3825-3831.
26. Robinson, C.K. et al. (2011). A major role for nonenzymatic antioxidant processes in the radioresistance of Halobacterium salinarum. J. Bacteriol. 193:1653-1662.
27. Dai, D.F. et al. (2011). Mitochondrial targeted antioxidant peptide ameliorates hypertensive cardiomyopathy. J. Am. Coll. Cardiol. 58:73-82.
28. Thompson, C.M. et al. (2011). Investigation of the mode of action underlying the tumorigenic response induced in B6C3F1 mice exposed orally to hexavalent chromium. Toxicol. Sci. 123:58-70.
29. Ungvari, Z. et al. (2011). Extreme longevity is associated with increased resistance to oxidative stress in Arctica islandica, the longest-living non-colonial animal. J. Gerontol A Biol Sci Med Sci. 10.1093/gerona/glr044
30. Labbé, A. et al. (2010). Resveratrol improves insulin resistance hyperglycemia and hepatosteatosis but not hypertriglyceridemia, inflammation, and life span in a mouse model for Werner Syndrome. J. Gerontol. A Biol Sci Med. Sci. 10.1093/gerona/glq184.
31. Nolin, T.D. et al (2010). Multiple-dose pharmacokinetics and pharmacodynamics of N-acetylcysteine in patients with end-stage renal disease. Clin. J. Am. Soc. Nephrol. 5:1588-1594.
32. Kang, K.A. et al. (2010). KIOM-4 protects against oxidative stress-induced mitochondrial damage in pancreatic beta-Cells via its antioxidant effects. Evid. Based Complement. Altern. Med. 10.1093/ecam/neq007.
33. Cho, H-Y. et al. (2009). Antiviral activity of Nrf2 in a murine model of respiratory syncytial virus (RSV) disease. Am. J. Respir. Crit. Care Med. 179:138-150.
34. Neretti, N. et al. (2009). Long lived Indy induces reduced mitochondrial reactive oxygen species production and oxidative damage. PNAS 106:2277-2282.
35. Maity, P. et al. (2008). Indomethacin, a non-steroidal anti-inflammatory drug, develops gastropathy by inducing reactive oxygen species-mediated mitochondrial pathology and associated apoptosis in gastric mucosa: a novel role of mitochondrial aconitase oxidation. J. Biol. Chem. 284:3058-3068.
36. Kim, S-H. et al. (2008). Protective effect of esculetin against oxidative stress-induced cell damage via scavenging reactive oxygen species. Acta Pharmacol. Sinica 29(11):1319-1326.
37. Paromov, V. et al. (2008). The influence of N-acetyl-L-cysteine on oxidative stress and nitric oxide synthesis in stimulated macrophages treated with a mustard gas analogue. BMC Cell Biology 9:33.
38. Lowry, M. et al. (2007). Lung lining fluid glutathione attenuates IL-13 induced asthma. Am. J. Respir. Cell Mol. Biol. 38(5):509-516.
Similar Products
Product Notes
The OxiSelect Protein Carbonyl (Catalog #AAA168972) is an ELISA Kit and is intended for research purposes only. The product is available for immediate purchase. It is sometimes possible for the material contained within the vial of "OxiSelect Protein Carbonyl, ELISA Kit" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
If you are ready to order, navigate to Shopping Cart and get ready to checkout.
