P6140
Peroxidasa from horseradish
Type X, ammonium sulfate suspension
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 X
form
ammonium sulfate suspension
specific activity
≥225 units/mg protein (biuret, using pyrogallol)
mol wt
~44 kDa
concentration
≥1.0 mg/mL
absorbance ratio
RZ 2.5-3.5
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. 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.
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 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.
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.
Physical form
Crystalline suspension in 3.2 M (NH4)2SO4 solution containing potassium phosphate buffer, pH 6.0
Preparation Note
Water may be used to dilute suspension if needed.
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.
Preliminary studies indicate the presence of two basic and no acidic isoenzymes
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
11 - 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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Artículos
Instructions for working with enzymes supplied as ammonium sulfate suspensions
Protocolos
Reinheitszahl (RZ) is the ratio of absorbance due to hemin (A403, Soret region) to absorbance due to protein (A275).
This procedure is for the determination of Peroxidase enzymatic activity using Pyrogallol as the substrate.
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