P6140
Peroxydase from horseradish
Type X, ammonium sulfate suspension
Synonyme(s) :
Donneur : peroxyde d'hydrogène oxydoréductase, Peroxydase de raifort
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About This Item
Produits recommandés
Source biologique
horseradish
Type
Type X
Forme
ammonium sulfate suspension
Activité spécifique
≥225 units/mg protein (biuret, using pyrogallol)
Poids mol.
~44 kDa
Concentration
≥1.0 mg/mL
Rapport des absorbances
RZ 2.5-3.5
Température de stockage
2-8°C
InChI
1S/H2O3/c1-3-2/h1-2H
Clé InChI
JSPLKZUTYZBBKA-UHFFFAOYSA-N
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Catégories apparentées
Description générale
La peroxydase de raifort (HRP, de l′anglais horseradish peroxidase) est isolée des racines de raifort (Armoracia rusticana) et appartient au groupe des peroxydases de type ferroprotoporphyrine. La HRP est un polypeptide à une seule chaîne qui comporte quatre ponts disulfure. Il s′agit d′une glycoprotéine contenant 18 % de glucides. Selon l′isozyme, les glucides peuvent inclure les constituants suivants : galactose, arabinose, xylose, fucose, mannose, mannosamine et galactosamine. Le poids moléculaire de la peroxydase de raifort (env. 44 kDa) inclut la chaîne polypeptidique (33,890 Daltons), l′hémine plus Ca2+ (env. 700 Daltons) et les glucides (env. 9,400 Daltons). Au moins sept isozymes existent. Le point isoélectrique des isozymes de peroxydase de raifort se situe dans une plage de 3,0 à 0,9.
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.
Actions biochimiques/physiologiques
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.
Définition de l'unité
Une unité de pyrogallol forme 1,0 mg de purpurogalline à partir de pyrogallol en 20 sec à pH 6,0 et 20 °C.
Forme physique
Crystalline suspension in 3.2 M (NH4)2SO4 solution containing potassium phosphate buffer, pH 6.0
Notes préparatoires
Water may be used to dilute suspension if needed.
Remarque sur l'analyse
La valeur RZ (Reinheitszahl) est le rapport d′absorbance A403/A275 mesuré à 0,5-1,0 mg/ml dans de l′eau désionisée. C′est une mesure de la teneur en hémine, plutôt que de l′activité enzymatique. Il est possible que des préparations présentant une valeur RZ élevée aient une faible activité enzymatique.
Preliminary studies indicate the presence of two basic and no acidic isoenzymes
Autres remarques
Pour en savoir plus sur la peroxydase, rendez-vous sur www.sigma-aldrich.com/enzymeexplorer.
Mention d'avertissement
Danger
Mentions de danger
Conseils de prudence
Classification des risques
Resp. Sens. 1
Code de la classe de stockage
11 - Combustible Solids
Classe de danger pour l'eau (WGK)
WGK 1
Équipement de protection individuelle
Eyeshields, Gloves, type N95 (US)
Certificats d'analyse (COA)
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Vascular permeability in various rat sympathetic ganglia, including superior cervical ganglia, thoracic ganglia and the celiac-mesenteric ganglia (CMG) complex, was investigated by using lanthanum and horseradish peroxidase (HRP) as tracers with special attention to the neuronal and small granule-containing (SGC)
Articles
Instructions for working with enzymes supplied as ammonium sulfate suspensions
Protocoles
Reinheitszahl (RZ) is the ratio of absorbance due to hemin (A403, Soret region) to absorbance due to protein (A275).
To standardize a procedure for the assay of Peroxidase using 2,2'-Azino-bis(3-Ethylbenzthiazoline-6-Sulfonic Acid) as a substrate.
This procedure is for the determination of Peroxidase enzymatic activity using Pyrogallol as the substrate.
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