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P6140

Sigma-Aldrich

Peroxidase from horseradish

Type X, ammonium sulfate suspension

Synonim(y):

Donor:hydrogen-peroxide oxidoreductase, Horseradish peroxidase

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About This Item

Numer CAS:
9003-99-0
Numer EC enzymu:
1.11.1.7 (BRENDA, IUBMB)
Numer WE:
232-668-6
Numer MDL:
MFCD00071339
Kod UNSPSC:
12352204
eCl@ss:
32160410
NACRES:
NA.54

pochodzenie biologiczne

horseradish

typ

Type X

Postać

ammonium sulfate suspension

aktywność właściwa

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

masa cząsteczkowa

~44 kDa

stężenie

≥1.0 mg/mL

stosunek absorbancji

RZ 2.5-3.5

temp. przechowywania

2-8°C

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. 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.

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 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.

Definicja jednostki

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

Postać fizyczna

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

Uwaga dotycząca przygotowania

Water may be used to dilute suspension if needed.

Komentarz do analizy

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.

Inne uwagi

View more information on peroxidase at www.sigma-aldrich.com/enzymeexplorer.
This page may contain text that has been machine translated.

inhibitor

Numer produktu
Opis
Cennik

substrat

Numer produktu
Opis
Cennik

Piktogramy

Health hazard
GHS08

Hasło ostrzegawcze

Danger

Zwroty wskazujące rodzaj zagrożenia

H334

Zwroty wskazujące środki ostrożności

P261 - P284 - P501

Klasyfikacja zagrożeń

Resp. Sens. 1

Kod klasy składowania

11 - Combustible Solids

Klasa zagrożenia wodnego (WGK)

WGK 1

Środki ochrony indywidualnej

Eyeshields, Gloves, type N95 (US)

Certyfikaty analizy (CoA)

Poszukaj Certyfikaty analizy (CoA), wpisując numer partii/serii produktów. Numery serii i partii można znaleźć na etykiecie produktu po słowach „seria” lub „partia”.

Masz już ten produkt?

Dokumenty związane z niedawno zakupionymi produktami zostały zamieszczone w Bibliotece dokumentów.

Odwiedź Bibliotekę dokumentów

David Stuart Thompson et al.
Plants (Basel, Switzerland), 10(7) (2021-07-03)
The extensibility of synthetic polymers is routinely modulated by the addition of lower molecular weight spacing molecules known as plasticizers, and there is some evidence that water may have similar effects on plant cell walls. Furthermore, it appears that changes
N Santanam et al.
Journal of lipid research, 39(11), 2111-2118 (1998-11-04)
Estradiol has been documented to inhibit the oxidation of low density lipoprotein (LDL). We show that physiological concentrations of estradiol do not inhibit the oxidation of LDL by copper. LDL samples isolated from a) premenopausal and postmenopausal women and from
Christoph Plieth et al.
Plant signaling & behavior, 7(6), 650-660 (2012-05-15)
In this paper we demonstrate how peroxidase (PO) activities and their heat stability correlate with the availability of free Ca(2+) ions. Calcium ions work as a molecular switch for PO activity and exert a protective function, rendering POs heat stable.
John W Callahan et al.
Methods in molecular biology (Clifton, N.J.), 347, 321-330 (2006-10-31)
This chapter describes in detail a practical procedure for the preparation of radiolabeled galactocerebroside and its use in the assay of galactocerebrosidase (GalCase), the enzyme deficient in globoid cell leukodystrophy (Krabbe disease). The reference range for leukocytes and fibroblasts is
G M Cook et al.
Microbiology (Reading, England), 144 ( Pt 12), 3297-3308 (1999-01-12)
Escherichia coli produces an extracellular factor that inhibits the aerobic growth of Cyd- mutants, defective in the synthesis or assembly of the cytochrome bd-type quinol oxidase. This paper shows that such a factor is the iron-chelating siderophore enterochelin. Mutants in
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Produkty

Horseradish Peroxidase (HRP) Enzymes

Discover our peroxidase from horseradish enzymes, products, substrates, and inhibitors for your ELISA, immunoassay, and protein application needs.

Enzymy peroksydazy chrzanowej (HRP)

Odkryj naszą peroksydazę z enzymów chrzanowych, produkty, substraty i inhibitory do testów ELISA, testów immunologicznych i zastosowań białkowych.

How to Work with Enzymes Supplied as Ammonium Sulfate Suspensions

Instructions for working with enzymes supplied as ammonium sulfate suspensions

Protokoły

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).

Enzymatic Assay of Peroxidase (EC 1.11.1.7) 2,2'-Azino-bis(3-Ethylbenzthiazoline-6-Sulfonic Acid) as a Substrate

To standardize a procedure for the assay of Peroxidase using 2,2'-Azino-bis(3-Ethylbenzthiazoline-6-Sulfonic Acid) as a substrate.

Enzymatic Assay of Peroxidase (EC 1.11.1.7)

This procedure is for the determination of Peroxidase enzymatic activity using Pyrogallol as the substrate.

Test enzymatyczny peroksydazy (EC 1.11.1.7)

Ta procedura służy do oznaczania aktywności enzymatycznej peroksydazy przy użyciu pirogallolu jako substratu.

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