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
biological source
horseradish
Quality Level
type
Type II
form
essentially salt-free, lyophilized powder
specific activity
150-250 units/mg solid (using pyrogallol)
mol wt
~44 kDa
solubility
0.1 M phosphate buffer: soluble (pH 6.0)
H2O: soluble
absorbance ratio
RZ ≥1.8
application(s)
diagnostic assay manufacturing
storage temp.
2-8°C
InChI
1S/H2O3/c1-3-2/h1-2H
InChI key
JSPLKZUTYZBBKA-UHFFFAOYSA-N
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General description
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.
Application
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.
Biochem/physiol Actions
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.
Quality
Preliminary work shows it to contain at least five isoenzymes.
Unit Definition
Una unidad de pirogalol formará 1,0 mg de purpurogalina a partir de pirogalol en 20 segundos a pH 6,0 a 20 °C.
Analysis Note
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.
Other Notes
Véase más información sobre la peroxidasa en www.sigma-aldrich.com/enzymeexplorer.
inhibitor
Referencia del producto
Descripción
Precios
substrate
Referencia del producto
Descripción
Precios
signalword
Danger
hcodes
pcodes
Hazard Classifications
Resp. Sens. 1
Storage Class
13 - Non Combustible Solids
wgk_germany
WGK 1
ppe
Eyeshields, Gloves, type N95 (US)
Certificados de análisis (COA)
Busque Certificados de análisis (COA) introduciendo el número de lote del producto. Los números de lote se encuentran en la etiqueta del producto después de las palabras «Lot» o «Batch»
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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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