POD Immunostain Set

for POD Staining

Manufacturer :: FUJIFILM Wako Pure Chemical Corporation

Storage Condition :: Keep at 2-10 degrees C. (RT)

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Comparison	Product Number	Package Size	Price	Availability	Certificate of Analysis	Purchase
	Distributor 299-18841 Barcode No 4987481359093	20mL x12	List Price 449.00 USD	In stock in Japan	Certificate of Analysis	Distributor

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Package Insert

20mL x12

Certificate of Analysis

20mL x12

Calibration Certificate

Analytical Charts

Overview / Applications

Outline	This product is for research use only. Do not administer it to human. Peroxidase has been widely employed as labeling enzyme in an enzyme-linked immunostain method by Western blotting for detection of slight amount of protein. However, diaminobenzidine (DAB) and 4-Chloro-1-naphtol which are the substrate of POD staining method has the problem of sensitivity, colorization and carcinogenic activity pointed out. POD Immunostain Set resolves this problem, allowing to include all the reagents required for staining. Involves simple preparation, and by an optimized tetrazolium method gives the greater color intensities. [Kit contents] (1) NTB solution (contains 0.02 % Phenol): 1 bottle x 250 mL (2) NADH: 12 bottles x for 20 mL (3) Substrate solution (Contains 0.02 % H₂O₂): 1 bottle x 13 mL [Staining principle] Phenol included in the kit is oxidized by POD and H2O2. In the presence of NADH and NTB, violet diformazan proportionated to POD activation is formed. [Staining preparation] Dissolve one bottle of NADH in 20 mL NTB Solution. Add 1 mL of Substrate Solution to the solution and made it as Staining Solution. After preparation, it is possible to use for five days at 2 ~ 10 degrees C in a dark place. [Staining procedure] Wash the support by a common method after POD labeling antibody reaction. Transfer the support to Staining Solution* and make it react for 10 minutes at the room temperature. Wash it to stop the reaction. The reaction time of 10 minutes is standard time, so by the amount of a sample adjust the time. It will become possible to preserve for long time after dried and stored in a dark place. * Note: The amount of Staining Solution is enough to cover the support but please make sure that there is no shortage of solution. [Technical notes] The staining of the background proceeds from long-time reaction. Reacting under the light shield reduces the staining of the background. We recommend to wash the tray immediately after the staining is finished because Staining Solution which is left in the tray for longtime causes to stain the tray itself. In immunohistochemistry, using xylene solution as an inclusion causes the color reagents to be dissolved. [References] Taketa, K., Ichikawa, E. and Hanada, T.: J. Immunol. Methods, 95, 71 (1986). Hokari, S., Hasegawa, M., Tanaka, M., Sakagishi, Y. and Kikuchi, G.: J. Biochem., 104, 211 (1988).

Outline

This product is for research use only. Do not administer it to human.

Peroxidase has been widely employed as labeling enzyme in an enzyme-linked immunostain method by Western blotting for detection of slight amount of protein. However, diaminobenzidine (DAB) and 4-Chloro-1-naphtol which are the substrate of POD staining method has the problem of sensitivity, colorization and carcinogenic activity pointed out.
POD Immunostain Set resolves this problem, allowing to include all the reagents required for staining. Involves simple preparation, and by an optimized tetrazolium method gives the greater color intensities.

[Kit contents]
(1) NTB solution (contains 0.02 % Phenol): 1 bottle x 250 mL
(2) NADH: 12 bottles x for 20 mL
(3) Substrate solution (Contains 0.02 % H₂O₂): 1 bottle x 13 mL

[Staining principle]
Phenol included in the kit is oxidized by POD and H2O2. In the presence of NADH and NTB, violet diformazan proportionated to POD activation is formed.
[Staining preparation]
Dissolve one bottle of NADH in 20 mL NTB Solution.
Add 1 mL of Substrate Solution to the solution and made it as Staining Solution. After preparation, it is possible to use for five days at 2 ~ 10 degrees C in a dark place.

[Staining procedure]

Wash the support by a common method after POD labeling antibody reaction.

Transfer the support to Staining Solution* and make it react for 10 minutes at the room temperature.

Wash it to stop the reaction. The reaction time of 10 minutes is standard time, so by the amount of a sample adjust the time.

It will become possible to preserve for long time after dried and stored in a dark place.

* Note: The amount of Staining Solution is enough to cover the support but please make sure that there is no shortage of solution.

[Technical notes]

The staining of the background proceeds from long-time reaction. Reacting under the light shield reduces the staining of the background.

We recommend to wash the tray immediately after the staining is finished because Staining Solution which is left in the tray for longtime causes to stain the tray itself.

In immunohistochemistry, using xylene solution as an inclusion causes the
color reagents to be dissolved.

[References]

Taketa, K., Ichikawa, E. and Hanada, T.: J. Immunol. Methods, 95, 71 (1986).

Hokari, S., Hasegawa, M., Tanaka, M., Sakagishi, Y. and Kikuchi, G.: J. Biochem., 104, 211 (1988).

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For research use or further manufacturing use only. Not for use in diagnostic procedures.

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