• USA Home
  • P8415 - Peroxidase from horseradish

EMAIL THIS PAGE TO A FRIEND
P8415 Sigma

Peroxidase from horseradish

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

Synonym: Donor:hydrogen-peroxide oxidoreductase, Horseradish peroxidase

Purchase

Description

Analysis Note

Preliminary isoelectric focusing data indicates this is primarily isoenzyme C

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

A further purification of Peroxidase TypeVI (P8375).

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

Packaging

500, 1000, 2000, 5000 units in glass bottle

Preparation Note

Chromatographically purified

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 P8415, type XII, is an essentially salt free lyophilized powder. It is a further purification of product P8375. It is commonly used to determine amounts of glucose and peroxides in solution. It has been used in an aspergillus fumigatus rapid susceptibility assay.

The enzyme has been used to determine H2O2 production in tobacco BY-2 cells using a spectrofluorimetric method.

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. It is also used for the determination of glucose and peroxides in solution.

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

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.

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

Suggested Laboratory Gloves


Laboratory GlovesThis substance has been tested against several types of hand protection for CE compliance. Click below to find the recommended gloves for handling this product.




All labs need water
Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
3

Documents

Certificate of Analysis

Certificate of Origin

Protocols & Articles

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

Set your institution to view full text papers.
Related Products

inhibitor

Product #

Description

Add to Cart

S2002 Sodium azide, ReagentPlus®, ≥99.5%

related product

Product #

Description

Add to Cart

08168 Timestrip Plus 8 °C

substrate

Product #

Description

Add to Cart

D9143 o-Dianisidine, peroxidase substrate
A0156 N-(4-Aminobutyl)-N-ethylisoluminol, ≥90%
A4685 Luminol sodium salt
A1888 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, ≥98% (HPLC)
G5502 Guaiacol, oxidation indicator
A1661 N-(6-Aminohexyl)-N-ethylisoluminol
A3537 5-Aminosalicylic acid, ≥99%
A4382 4-Aminoantipyrine, reagent grade
A8264 4-Aminophthalhydrazide
A5754 3-Amino-9-ethylcarbazole, ≥95%, powder
A8511 Luminol, ≥97% (HPLC)

Technical Service:

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

Bulk Ordering & Pricing:

Need larger quantities for your development, manufacturing or research applications?