Merck
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P6140

Sigma-Aldrich

Peroxidase from horseradish

Type X, ammonium sulfate suspension

Synonym(s):
Donor:hydrogen-peroxide oxidoreductase, Horseradish peroxidase
CAS Number:
Enzyme Commission number:
Número EC:
Número MDL:
eCl@ss:
32160410
NACRES:
NA.54

fonte biológica

horseradish

Nível de qualidade

200

tipo

Type X

forma

ammonium sulfate suspension

specific activity

≥225 units/mg protein (biuret, using pyrogallol)

peso molecular

~44 kDa

concentração

≥1.0 mg/mL

razão de absorbância

RZ 2.5-3.5

temperatura de armazenamento

2-8°C

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Descrição geral

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). 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 P6140 has been used to detect low density lipoprotein (LDL).
The enzyme from Sigma has been used while assessing the skin sensitization potential of pro-haptens. It has also been used to show that peroxidase (PO) activity and its heat stability correlate with the availability of free Ca2+ ions.

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 that include the use of glutaraldehyde, periodate oxidation, 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. 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. Sodium azide, cyanide, L-cystine, dichromate, ethylenethiourea, hydroxylamine, sulfide, vanadate, p-aminobenzoic acid, and Cd2+, Co2+, Cu2+, Fe3+, Mn2+, Ni2+, Pb2+ ions are found to inhibit its enzyme activity.
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.

Definição da unidade

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

forma física

Crystalline suspension in 3.2 M (NH4)2SO4 solution containing potassium phosphate buffer, pH 6.0

Nota de preparo

Water may be used to dilute suspension if needed.

Nota de análise

Preliminary studies indicate the presence of two basic and no acidic isoenzymes
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.

Outras notas

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

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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