P8125

Peroxidasa from horseradish

Type I, essentially salt-free, lyophilized powder, ≥50 units/mg solid (using pyrogallol)

• Sinónimos: Donante:peróxido de hidrógeno oxidorreductasa, Peroxidasa de rábano picante
• Número de CAS: 9003-99-0
• Comisión internacional de enzimas: 1.11.1.7 (BRENDA, IUBMB)
• EC Number: 232-668-6
• MDL number: MFCD00071339
• UNSPSC Code: 12352204
• eCl@ss: 32160410
• NACRES: NA.54

Propiedades

• type: Type I
• Quality Level: 200
• form: essentially salt-free, lyophilized powder
• specific activity: ≥50 units/mg solid (using pyrogallol)
• mol wt: ~44 kDa
• solubility: 0.1 M phosphate buffer: soluble (pH 6.0); H₂O: soluble
• absorbance ratio: RZ ≥1.0
• storage temp.: 2-8°C
• InChI: 1S/H2O3/c1-3-2/h1-2H
• Inchi Key: JSPLKZUTYZBBKA-UHFFFAOYSA-N

Descripción

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

Peroxidase from horseradish has been used:

• in fecal starch analysis
• to measure plasma glucose levels by Trinder method
• to incubate feruloylated arabinoxylan fractions and also to evaluate oxidative coupling
• in the detection of glycoproteins

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 known 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.

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.

Información de seguridad

• pictograms: GHS08
• signalword: Danger
• hcodes: H334
• pcodes: P261 - P284 - P501
• Hazard Classifications: Resp. Sens. 1
• Storage Class: 11 - Combustible Solids
• wgk_germany: WGK 1
• ppe: Eyeshields, Gloves, type N95 (US)