P8250
Peroxidasa from horseradish
Type II, essentially salt-free, lyophilized powder, 150-250 units/mg solid (using pyrogallol)
Sinónimos:
Donante:peróxido de hidrógeno oxidorreductasa, Peroxidasa de rábano picante
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About This Item
Productos recomendados
origen biológico
horseradish
Nivel de calidad
tpo
Type II
Formulario
essentially salt-free, lyophilized powder
actividad específica
150-250 units/mg solid (using pyrogallol)
mol peso
~44 kDa
solubilidad
0.1 M phosphate buffer: soluble (pH 6.0)
H2O: soluble
cociente de absorbancia
RZ ≥1.8
aplicaciones
diagnostic assay manufacturing
temp. de almacenamiento
2-8°C
cadena SMILES
[O+H2]O[O-]
InChI
1S/H2O3/c1-3-2/h1-2H
Clave InChI
JSPLKZUTYZBBKA-UHFFFAOYSA-N
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Descripción general
La peroxidasa de rábano picante se aísla de las raíces de rábano picante (Amoracia rusticana) y pertenece al grupo de ferroprotoporfirina de las peroxidasas. La HRP es un polipéptido monocatenario que contiene cuatro puentes bisulfuro. Es una glucoproteína que contiene un 18 % de carbohidratos. La composición de los carbohidratos consiste en galactosa, arabinosa, xilosa, fucosa, manosa, manosamina y galactosamina dependiendo de la isoenzima específica. Su peso molecular (~44 kDa) incluye la cadena polipeptídica (33 890 dalton), la hemina más Ca2+ (~700 dalton) y los carbohidratos (~9 400 dalton). Existen al menos siete isoenzimas de la HRP. El punto isoeléctrico de las isoenzimas de la peroxidasa de rábano picante oscila entre 3,0 y 9,0.
Aplicación
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.
Acciones bioquímicas o 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. 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.
Calidad
Preliminary work shows it to contain at least five isoenzymes.
Definición de unidad
Una unidad de pirogalol formará 1,0 mg de purpurogalina a partir de pirogalol en 20 segundos a pH 6,0 a 20 °C.
Nota de análisis
La RZ (Reinheitszahl) es la relación de absorbancia A403/A275 determinada a 0,5-1,0 mg/ml en agua desionizada. Es una medida del contenido de hemina, no de la actividad enzimática. Incluso preparaciones con RZ elevada pueden tener una actividad enzimática baja.
Otras notas
Véase más información sobre la peroxidasa en www.sigma-aldrich.com/enzymeexplorer.
Inhibidor
Referencia del producto
Descripción
Precios
sustrato
Referencia del producto
Descripción
Precios
Palabra de señalización
Danger
Frases de peligro
Consejos de prudencia
Clasificaciones de peligro
Resp. Sens. 1
Código de clase de almacenamiento
13 - Non Combustible Solids
Clase de riesgo para el agua (WGK)
WGK 1
Equipo de protección personal
Eyeshields, Gloves, type N95 (US)
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Protocolos
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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