Test kit for Potential Anti Oxidant (PAO)

Catalog No. ABIN1020372

Product Details

Reactivity: Human

Detection Range: 21.9-4378 μmol/L

Minimum Detection Limit: 21.9 μmol/L

Purpose: Detection: 480-492 nm, colorimetric assay.
In the PAO assay kit, an easy and convenient method to measure antioxidant capacity is provided. Utilizing the reduction of cupric ion (Cu++ to 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, beverage

Sample Type: Serum, Food, Beverages

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

Material not included: 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).

Application Details

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 µL of Sample Diluent, and add 10 µL of standards or diluted samples.
3) Pour 200 µL of mixture to Micro titer plate. Use 200 µL of Sample Diluent for blank well.
4) Read absorbance at 490 nm (as READ1).
5) Add 50 µL of Cu++solution to each well, mix gently, and incubate at room temperature for 3 minutes.
6) Add 50 µL 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 (µmol/L). 1mM of uric acid = 2189 µmol/L (copper reducing power)

Restrictions: For Research Use only

Handling

Storage: RT

References

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Kadowaki, Anraku, Tasaki, Taguchi, Shimoishi, Seo, Hirata, Maruyama, Otagiri: "Evaluation for antioxidant and renoprotective activity of olmesartan using nephrectomy rats." in: Biological & pharmaceutical bulletin, Vol. 32, Issue 12, pp. 2041-5, (2009) (PubMed).

Yokohira, Yamakawa, Saoo, Matsuda, Hosokawa, Hashimoto, Kuno, Imaida: "Antioxidant effects of flavonoids used as food additives (purple corn color, enzymatically modified isoquercitrin, and isoquercitrin) on liver carcinogenesis in a rat medium-term bioassay." in: Journal of food science, Vol. 73, Issue 7, pp. C561-8, (2008) (PubMed).

Straface, Matarrese, Gambardella, Vona, Sgadari, Silveri, Malorni: "Oxidative imbalance and cathepsin D changes as peripheral blood biomarkers of Alzheimer disease: a pilot study." in: FEBS letters, Vol. 579, Issue 13, pp. 2759-66, (2005) (PubMed).

Pregel, Bollo, Cannizzo, Biolatti, Contato, Biolatti: "Antioxidant capacity as a reliable marker of stress in dairy calves transported by road." in: The Veterinary record, Vol. 156, Issue 2, pp. 53-4, (2005) (PubMed).

Vassalle, Petrozzi, Botto, Andreassi, Zucchelli: "Oxidative stress and its association with coronary artery disease and different atherogenic risk factors." in: Journal of internal medicine, Vol. 256, Issue 4, pp. 308-15, (2004) (PubMed).

Target Details

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.