Genox

Potential Antioxidant Test Kit
Catalog #KPAO

Product Insert (pdf)


Antioxidant assay:

Oxidative stress plays an 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 Potential Antioxidant Test Kit (PAO), 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++ ions are reduced by antioxidants to form Cu+. Reduced Cu+ ions react with Chromatic Solution (Bathocuproine) , and can be detected by absorbance at wavelength 480 to 490 nm. Antioxidant capacity can be calculated from the amount of Cu+ ions 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.

Specifications

Assay principle:

Colorimetric  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).

Assay principle:

Colormetric assay(detection: 480 - 492 nm)

Assay range:

21.9 - 4378 micro mol/L (cupric ion reducing power)

Format:

96 wells

 

Content of this kit

Standard (Uric acid powder):

1 vial

Sample diluent:

2 bottles

Cu++ solution:

1 bottle

Stop solution:

1 bottle

Micro titer plate:

1 plate (96 wells)

 

 

POA Test Kit 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)

 


REFERENCES

1)

Oxidative imbalance and cathepsin D changes as peripheral blood biomarkers of Alzheimer disease: A pilot study.
E Strafacea, P Matarresea, L Gambardella, R Vona, A Sgadari,MC Silveri, W Malorni
FEBS Letters 579, p2759-766 (2005)

2)

Oxidative stress and its association with coronary artery disease and different atherogenic risk factors.
C. VASSALLE, L. PETROZZI , N. BOTTO, M. G. ANDREASSI and G. C. ZUCCHELLI
Journal of Internal Medicine 256: 308-315 (2004)

3)

Antioxidant capacity as a reliable marker of stress in dairy calves transported by road.
P Pregel, E Bollo, FT Cannizzo, B Biolatti, E Contato, and PG Biolatti
Veterinary Record 156, p53-54 (2005)

4)

Vitamin E-coated dialyzers reduce oxidative stress related proteins and markers in hemodialysis ? a molecular biological approach.
LA Calo, A Naso, E Pagnin, PA Davis, M Castoro, R Corradin, P Riegler, C Cascone, W Huber and A Piccoli
Clinical Nephrology, Vol.62(5), p355-361 (2004)

5)

Oxidative stress-related factors in Bartter's and Gitelman9s syndrome: relevance for angiotensin II signalling.
Calo LA, Pagnin E, Davis PA, Sartori M, Semplicini A.
Nephrol Dial Transplant ,Vol.18(8) p1518-25 (2003)

6)

Effect of epoetin on HO-1 mRNA level and plasma antioxidants in hemodialysis patients.
Calo LA, Stanic L, Davis PA, Pagnin E, Munaretto G, Fusaro M, Landini S, Semplicini A, Piccoli A.
Int. J Clin. Ther, Vol.41(5), p187-92 (2003)

7)

Restored Antioxidant Capacity Parallels the Immunologic and Virologic Improvement in Children with Perinatal Human Immunodeficiency Virus Infection Receiving Highly Active Antiretroviral Therapy.
M Martino, F Chiarelli, M Moriondo, M Torello, C Azzari, and L Galli
Clinical Immunology, Vol.100(1),p82-6 (2001)
The ‘Potential Antioxidant Test Kit’ is manufactured in Japan by the Japan Institute for the Control of Aging (JaICA). Genox is the distributor of this kit for users in North and South America.

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Note: Genox’s Products are for RESEARCH USE ONLY.
Users of Genox’s products are strongly advised NOT TO USE THEM FOR CLINICAL/DIAGNOSTIC APPLICATIONS. 

The provision of test reports, generated by using Genox's products, to individuals
or to treating physicians for the diagnosis, prevention, treatment and control of any human disease or impairment of, or the assessment of the health, nutritional, or medical condition of individuals is expressly prohibited by law.  42 U.S.C. § 263a (2006); 42 C.F.R. § 493.3 (2006); COMAR 10.10.01.02 (2006). Genox is not a CLIA certified laboratory.

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