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Ready-to-use Substrate Solution for Peroxidase

Peroxidase is an enzyme that catalyzes the oxidation of a given substrate mediated by hydrogen peroxide and is often used in biochemical experiments. 1 and 2 are available as follows.

1 is supplied as a ready-to-use solution for ELISA containing both 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid ammonium salt) (AzBTS) and hydrogen peroxide. When 1 reacts with horseradish peroxidase (HRP), a green soluble reaction product appears. The absorbance of the reaction product is measured at 405 nm.
Direction for use
The following procedure is for a 96-well plate format.
1. Bring 1 to room temperature and mix gently.
2. Add 100 μL of 1 to each well.
3. Incubate the plate at room temperature for 30 minutes.
4. Within 1 hour from the start of the reaction, measure the absorbance of each well at 405 nm.

Figure 1. Example of use


2 is supplied as a ready-to-use solution containing both 4-CN (4-chloro-1-naphthol) and hydrogen peroxide for Western blotting. When 2 reacts with horseradish peroxidase (HRP), a blue-purple precipitate appears.
Direction for use
1. Incubate a blotting membrane with an HRP-conjugated antibody and then wash the membrane.
2. Incubate the washed membrane with 2 until color develops.
3. Dispose the solution and add deionized water to stop color development.

Firure 2. Example of Western blotting


These products are for research purpose only.

References

  • 1)Antibodies, A Laboratory Manual, ed. by E. Harlow and D. Lane. Cold Spring Harbor Laboratory Press, New York, 1988, p. 404, p. 506, p. 594.
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