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P8375 Sigma

Peroxidase from horseradish

Type VI, essentially salt-free, lyophilized powder, ≥250 units/mg solid (using pyrogallol)

Synonym: Donor:hydrogen-peroxide oxidoreductase, Horseradish peroxidase

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Description

Frequently Asked Questions

Frequently Asked Questions are available for this Product.

Analysis Note

The RZ (Reinheitszahl) is the absorbance ratio A403/A275 determined at 0.5-1.0 mg/ml in deionized water. It is a measure of hemin content, not enzymatic activity. Even preparations with high RZ may have low enzymatic activity.

Other Notes

View more information on peroxidase at www.sigma-aldrich.com/enzymeexplorer.

Packaging

1000, 2000 units in glass bottle

5000 units in poly bottle

10000, 25000, 100000, 250000 units in glass bottle

Unit Definition

One pyrogallol unit will form 1.0 mg purpurogallin from pyrogallol in 20 sec at pH 6.0 at 20 °C.

Application

Horseradish peroxidase (HRP) is isolated from horseradish roots (Amoracia rusticana) and belongs to the ferroprotoporphyrin group of peroxidases.. It is used in biochemistry applications such as western blots, ELISA and Immunohistochemistry. Horseradish peroxidase is used to amplify a weak signal and increase detectability of a target molecule, such as a protein. Product P8375, type VI, is an essentially salt free lyophilized powder. It is commonly used to determine amounts of glucose and peroxides in solution. It has been used to study sensory input of cervical spinal cord neurons.

The enzyme has been used to develop a thermostable soybean peroxidase-based biosensor by cross-linking and electrically ′wiring′ the enzyme through a redox-conducting hydrogel to a glassy carbon electrode. It has also been used to assay the oxidation of low density lipoprotein.

Biochem/physiol Actions

HRP readily combines with hydrogen peroxide (H2O2) and the resultant [HRP-H2O2] complex can oxidize a wide variety of hydrogen donors. The optimal pH is 6.0-6.5 and the enzyme is most stable in the pH range of 5.0-9.0. HRP can be conjugated to antibodies by several different methods including glutaraldehyde, periodate oxidation, through disulfide bonds, and also via amino and thiol directed cross-linkers. It is smaller and more stable than the enzyme labels β-galactosidase and alkaline phosphatase and hence, it is the most desired label. Also, its glycosylation leads to lower non-specific binding.

When incubated with a substrate, horseradish peroxidase produces a coloured, fluorimetric, or luminescent derivative of the labeled molecule, allowing quantification. Horseradish peroxidase has been shown to slightly reduce the level of inhibition in a cydAB mutant. Known inhibitors are sodium azide, cyanide, L-cystine, dichromate, ethylenethiourea, hydroxylamine, sulfide, vanadate, p-aminobenzoic acid, and Cd2+, Co2+, Cu2+, Fe3+, Mn2+, Ni2+, and Pb2+ ions.

General description

Horseradish peroxidase is isolated from horseradish roots (Amoracia rusticana) and belongs to the ferroprotoporphyrin group of peroxidases. HRP is a single chain polypeptide containing four disulfide bridges. It is a glycoprotein containing 18% carbohydrate. The carbohydrate composition consists of galactose, arabinose, xylose, fucose, mannose, mannosamine, and galactosamine depending upon the specific isozyme. Its molecular weight (~44 kDa) includes the polypeptide chain (33,890 Daltons), hemin plus Ca2+ (~700 Daltons), and carbohydrate (~9,400 Daltons). At least seven isozymes of HRP exist. The isoelectric point for horseradish peroxidase isozymes ranges from 3.0 - 9.0.

Price and Availability


Amplified Detection

All labs need water

Biomedical Applications
Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
3

Frequently Asked Questions

Which document(s) contains shelf-life or expiration date information for a given product?
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis. These documents are located on the product detail page under Useful Links & Tools. Click on the following link to search for a Certificate of Analysis. Please click the following link to see the details on our Product Dating Information.
How do I get lot-specific information or a Certificate of Analysis?
A Certificate of Analysis is available by lot number and can be obtained through our Advanced Search Option: http://www.sigmaaldrich.com/catalog/AdvancedSearchPage.do
How do I convert units of Peroxidase from horseradish, Product P8375, to solid?
The activity listed on the Certificate of Analysis for each lot can be used to convert from units to milligrams of solid.
What is the optimal pH range of Peroxidase from horseradish, Product P8375?
The optimal pH range for product P8375 is 6.0 - 6.5 as stated on our data sheet (http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/datasheet2/p8375dat.Par.0001.File.tmp/p8375dat.pdf).
What can you tell me about the stability of Peroxidase from horseradish, Product P8375, (as is, in suspension and in solution)?
This product is stable as a powder, kept refrigerated. See the certificate of analysis for a lot-specific recommended retest date. It is also stable as a crystalline suspension of 3.2 M (NH4)2SO4 solution containing potassium phosphate buffer, pH 6.0 (see Product No. P6140). It is reasonably stable in solution in phosphate buffer. We have found that at 10 mg/mL in 0.1 phosphate buffer, pH 6.0, solutions kept frozen at -20°C lose less than 2% of their activity per week.
How do I find price and availability?
There are several ways to find pricing and availability for our products.  Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote.  USA customers:  1-800-325-3010 or view local office numbers. 
What is the Department of Transportation shipping information for this product?
Transportation information can be found in Section 14 of the product's (M)SDS. To access the shipping information for this material, use the link on the product detail page for the product, or search here. 
My question is not addressed here, how can I contact Technical Service for assistance?
Use the option to the right to "Ask a Question" by email of a Technical Service Scientist.
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Protocols & Articles

Articles

Purification of Peroxidase by HPLC on Concanavalin A Coupled to TSKgel® Tresyl-5PW

From our library of Articles, Sigma-Aldrich presents Purification of Peroxidase by HPLC on Concanavalin A Coupled to TSKgel® Tresyl-5PW
Keywords: Chromatography, High performance liquid chromatography, Ligands

Protocols

Enzymatic Assay of Peroxidase (EC 1.11.1.7)

This procedure is for the determination of Peroxidase enzymatic activity using Pyrogallol as the substrate. Products using this method include, but are not limited to, P1709, P2088, P6140, P6782, P81...
Keywords: Enzyme activity, Extinction coefficient

Spectrophotometric Determination of Reinheitszahl (RZ) for Peroxidase (EC 1.11.1.7)

Reinheitszahl (RZ) is the ratio of absorbance due to hemin (A403, Soret region) to absorbance due to protein (A275). This ratio (A403/A275) is not a measure of enzyme activity. Products using this me...
Keywords: Enzyme activity

Related Content

Enzymes & Proteins

Application Index | Enzyme Index | Substrate Index | Inhibitor Index | Cofactor Index | Lectin Index
Keywords: Cell signaling, Diagnostic, Drug discovery, Molecular biology

Peer-Reviewed Papers
15

References

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