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P8250
Peroxidase from horseradish
Type II, essentially salt-free, lyophilized powder, 150-250 units/mg solid (using pyrogallol)
Synonim(y):
Donor:hydrogen-peroxide oxidoreductase, Horseradish peroxidase
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About This Item
Polecane produkty
pochodzenie biologiczne
horseradish
Poziom jakości
typ
Type II
Formularz
essentially salt-free, lyophilized powder
aktywność właściwa
150-250 units/mg solid (using pyrogallol)
masa cząsteczkowa
~44 kDa
rozpuszczalność
0.1 M phosphate buffer: soluble (pH 6.0)
H2O: soluble
stosunek absorbancji
RZ ≥1.8
Zastosowanie
diagnostic assay manufacturing
temp. przechowywania
2-8°C
ciąg SMILES
[O+H2]O[O-]
InChI
1S/H2O3/c1-3-2/h1-2H
Klucz InChI
JSPLKZUTYZBBKA-UHFFFAOYSA-N
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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
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. Horseradish peroxidase, product P8250, has been used to study nonoral antigens in inflamed gingiva and Ebola virus glycoprotein toxicity.
The enzyme has been used as a comparison during the peroxidase assay of extract from mature tall fescue leaf blades. It has also been used to measure H2O2 production.
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 found 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. Horseradish peroxidase has been shown to slightly reduce the level of inhibition in a cydAB mutant.
Jakość
Preliminary work shows it to contain at least five isoenzymes.
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.
Inne uwagi
View more information on peroxidase at www.sigma-aldrich.com/enzymeexplorer.
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Inhibitor
Numer produktu
Opis
Cennik
substrat
Numer produktu
Opis
Cennik
Hasło ostrzegawcze
Danger
Zwroty wskazujące rodzaj zagrożenia
Zwroty wskazujące środki ostrożności
Klasyfikacja zagrożeń
Resp. Sens. 1
Kod klasy składowania
13 - Non Combustible Solids
Klasa zagrożenia wodnego (WGK)
WGK 1
Środki ochrony indywidualnej
Eyeshields, Gloves, type N95 (US)
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Produkty
Odkryj naszą peroksydazę z enzymów chrzanowych, produkty, substraty i inhibitory do testów ELISA, testów immunologicznych i zastosowań białkowych.
Protokoły
Enzymatic Assay of Diamine Oxidase (E.C. No. 1.4.3.22)
This procedure may be used for all Alcohol Oxidase products.
Enzymatic Assay of Cholesterol Oxidase
Reinheitszahl (RZ) is the ratio of absorbance due to hemin (A403, Soret region) to absorbance due to protein (A275).
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