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For Oxidative Stress Research

Potential Antioxidant (PAO) assay Kit

   Oxidative stress marker product TOP

Background

Oxidative stress plays on important role in various diseases and aging. The control of oxidative stress is expected to be useful to prevent diseases and aging.Oxidative stress is caused by the imbalance between reactive oxygen species (ROS) and antioxidant defense system. For accurate assessment of oxidative stress, measurement of ROS, oxidative damage and antioxidant activity may be essential. Recently, antioxidants as functional foods which scavenge ROS attract a great deal of attention.
In the PAO assay kit, an easy and convenient method to measure antioxidant capacity is provided. Utilizing the reduction of cupric ion (Cu++?Cu+), antioxidant capacity of samples can be detected in 5 minutes. Samples are mixed with Cu++ Solution. Cu++ are reduced by antioxidants to form Cu+. Reduced Cu+ react with Chromatic Solution (Bathocuproine) , and can be detected by absorbance at wavelength 480 to 490 nm. Antioxidant capacity can be calculated from the Cu+ formed. PAO can detect not only hydrophilic antioxidants such as Vitamin C, glutathione, but also can detect hydrophobic antioxidants such as Vitamin E. Applicable for assessment of total antioxidants of serum, foods and beverage samples.

Features

Assay principle: Colormetric assay(detection: 480 - 492 nm)
Assay range: 21.9 - 4378 micro mol/L (cupric ion reducing power)
Format: 96 wells
Storage: Room temperature (10 - 25°C)
Applications:

Human and animal serum samples, foods and beverage samples.

 

Required but not provided: A micro plate reader (measuring wavelength 492 nm)
Pipettes and pipette chips
Plastic test tubes
Distilled water
NaOH, HCl solution and pH meter (Not required if standards are prepared with distilled water only).
Source Mouse
Immunogen 4-HNE-modified Keyhole Limpet Hemocyanin
Subclass IgG1(kappa)
Clone HNEJ-2
Form Lyophilized Powder
Specificity This antibody show almost negligible reactivity with proteins that were treated with other aldehydes, such as 2-nonenal, 2-hexenal, 1-hexanal, 4-hydroxy-2-hexenal,formaldehyde, or glutaraldehyde.
Applications Immunohistochemistry and Western blotting.
Storage Less than -20°C for 5 years

Content

Standard (Uric acid powder): 1 vial
Sample diluent: 1 bottles
Cu++ solution: 1 bottle
Stop solution: 1 bottle
Micro titer plate: 1 plate (96 wells)

Assay procedure

1) Prepare 6 levels of standards by diluting 2mM uric acid.
2) Please prepare plastic test tubes for 6 levels of standards and each sample. Pour 390 micro L of Sample Diluent, and add 10 micro L of standards or diluted samples.
3) Pour 200micro L of mixture to Micro titer plate. Use 200 microL of Sample Diluent for blank well.
4) Read absorbance at 490 nm (as READ1).
5) Add 50 micro L of Cu++solution to each well, mix gently, and incubate at room temperature for 3 minutes.
6) Add 50 micro L of Stop solution, mix gently, and read absorbance at 490 nm (as READ2).
7) Please draw standard curves by plotting the difference of absorbance readings (READ2 - READ1) as vertical axis, and concentration of uric acid standards (mM) as horizontal axis. Calculate the corresponding uric acid concentration of samples. Multiply corresponding uric acid concentration (mM) of samples by 2189, to estimate antioxidant power (micro mol/L).
1mM of uric acid = 2189 micro mol/L (copper reducing power)

Oxidative Stress Markers flyer download [PDF]

Product List

Product Name Cat# Quantity Price

Potential Anti Oxidant (PAO) kit

NNS-KPA-050E-EX 96WELL

¥ 89,600
$ 1195
€ 896

References
  •  Effetcts of the daily administration of a rehydrating supplement to trotter horses A Falaschini, G Marangoni, S Rizzi and MF Trombetta J Equine Sci 16(1),p1-9 (2005)
  •  Straface E. et al,;  Oxidative imbalance and cathepsin D changes as peripheral blood biomarkers of Alzheimer disease: a pilot study. FEBS Lett. 2005 May 23;579(13):2759-66. Epub 2005 Apr 19. PMID: 15907478
  •  Vassalle C. et al,;  Oxidative stress and its association with coronary artery disease and different atherogenic risk factors. J Intern Med. 2004 Oct;256(4):308-15. PMID: 15367173
  •  Pregel P. et al,;  Antioxidant capacity as a reliable marker of stress in dairy calves transported by road. Vet Rec. 2005 Jan 8;156(2):53-4. PMID: 15675527
  •  Calo LA. et al,;  Vitamin E-coated dialyzers reduce oxidative stress related proteins and markers in hemodialysis--a molecular biological approach. Clin Nephrol. 2004 Nov;62(5):355-61. PMID: 15571180
  •  Calo LA. et al,;  Oxidative stress-related factors in Bartter's and Gitelman's syndromes: relevance for angiotensin II signalling. Nephrol Dial Transplant. 2003 Aug;18(8):1518-25. PMID: 12897089
  •  Calo LA. et al,;  Effect of epoetin on HO-1 mRNA level and plasma antioxidants in hemodialysis patients. Int J Clin Pharmacol Ther. 2003 May;41(5):187-92. PMID: 12776808
  •  de Martino M. et al,;  Restored antioxidant capacity parallels the immunologic and virologic improvement in children with perinatal human immunodeficiency virus infection receiving highly active antiretroviral therapy. Clin Immunol. 2001 Jul;100(1):82-6. PMID: 11414748