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Potential Antioxidant(PAO) assay Kit
Suitable for detection of total antioxidant capacity in serum and food extracts. For research use only.

Antioxidant assay:

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.

Principle of this 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 and beverage samples.

 

Specifications:

Method: 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).

Content of this kit:
 
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)

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)

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