Merck
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P8415

Sigma-Aldrich

Peroxidase from horseradish

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

Synonym(s):
Donor:hydrogen-peroxide oxidoreductase, Horseradish peroxidase
CAS Number:
Enzyme Commission number:
Número EC:
Número MDL:
NACRES:
NA.54

tipo

Type XII

Nível de qualidade

200

forma

essentially salt-free, lyophilized powder

specific activity

≥250 units/mg solid (using pyrogallol)

peso molecular

~44 kDa

solubilidade

0.1 M phosphate buffer: soluble (pH 6.0)
H2O: soluble

razão de absorbância

RZ ≥3.0

temperatura de armazenamento

2-8°C

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Descrição geral

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.

Aplicação

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.
Peroxidase from horseradish has been used to initiate peroxidase-coupled assay. It has also been used in the preparation of β-galactosidase (β-gal) stock solution.
The enzyme has been used to determine H2O2 production in tobacco BY-2 cells using a spectrofluorimetric method.

Embalagem

500, 1000, 2000, 5000 units in glass bottle

Ações bioquímicas/fisiológicas

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.

Outras notas

A further purification of Peroxidase TypeVI (P8375).
View more information on peroxidase at www.sigma-aldrich.com/enzymeexplorer.

Definição da unidade

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

Nota de preparo

Chromatographically purified

Nota de análise

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.

Pictogramas

Health hazard

Palavra indicadora

Danger

Frases de perigo

Classificações de perigo

Resp. Sens. 1 - Skin Sens. 1

Código de classe de armazenamento

11 - Combustible Solids

WGK

WGK 3

Equipamento de proteção individual

Eyeshields, Gloves, type N95 (US)

Certificate of Analysis

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Certificate of Origin

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