P8125

Peroxidase from horseradish

Name: Peroxidase from horseradish
Brand: SIGMA

Type I, essentially salt-free, lyophilized powder, ≥50 units/mg solid (using pyrogallol)

Synonim(y):

Donor:hydrogen-peroxide oxidoreductase, Horseradish peroxidase

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About This Item

CAS Number:

9003-99-0

Numer WE:

1.11.1.7 (BRENDA, IUBMB)

Numer WE:

232-668-6

Numer MDL:

MFCD00071339

Kod UNSPSC:

12352204

eCl@ss:

32160410

NACRES:

NA.54

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typ

Type I

Poziom jakości

200

Formularz

essentially salt-free, lyophilized powder

aktywność właściwa

≥50 units/mg solid (using pyrogallol)

masa cząsteczkowa

~44 kDa

rozpuszczalność

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

stosunek absorbancji

RZ ≥1.0

temp. przechowywania

2-8°C

ciąg SMILES

[O+H2]O[O-]

InChI

1S/H2O3/c1-3-2/h1-2H

Klucz InChI

JSPLKZUTYZBBKA-UHFFFAOYSA-N

Opis ogólny

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.

Zastosowanie

Peroxidase from horseradish has been used:

in fecal starch analysis
to measure plasma glucose levels by Trinder method
to incubate feruloylated arabinoxylan fractions and also to evaluate oxidative coupling
in the detection of glycoproteins

Działania biochem./fizjol.

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.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 known to inhibit the enzyme activity.

When incubated with a substrate, horseradish peroxidase produces a coloured, fluorimetric, or luminescent derivative of the labeled molecule, allowing quantification.

Definicja jednostki

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

Komentarz do analizy

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.