P8125-25KU Display Image

Peroxidase from horseradish, Type I, essentially salt-free, lyophilized powder, 50-150 units/mg solid (using pyrogallol)

Code: P8125-25KU D2-231

Analysis Note

The RZ (Reinheitszahl) is the absorbance ratio A403/A275 determined at 0.5-1.0 mg/ml in deionized water. It is a measure of hemin content,...


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£157.00 EACH

Analysis Note

The RZ (Reinheitszahl) is the absorbance ratio A403/A275 determined at 0.5-1.0 mg/ml in deionized water. It is a measure of hemin content, not enzymatic activity. Even preparations with high RZ may have low enzymatic activity.

Application

Peroxidase from horseradish has been used:in fecal starch analysisto measure plasma glucose levels by Trinder methodto incubate feruloylated arabinoxylan fractions and also to evaluate oxidative couplingin the detection ofglycoproteins

Biochem/physiol Actions

When incubated with a substrate, horseradish peroxidase produces a coloured, fluorimetric, or luminescent derivative of the labeled molecule, allowing quantification.

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.

General description

Horseradish peroxidase is isolated from horseradish roots (Amoracia rusticana) and belongs to the ferroprotoporphyrin group of peroxidases. HRP is a single chain polypeptide containing four disulfide bridges. It is a glycoprotein containing 18% carbohydrate. The carbohydrate composition consists of galactose, arabinose, xylose, fucose, mannose, mannosamine, and galactosamine depending upon the specific isozyme. Its molecular weight (~44 kDa) includes the polypeptide chain (33,890 Daltons), hemin plus Ca2+ (~700 Daltons), and carbohydrate (~9,400 Daltons). At least seven isozymes of HRP exist. The isoelectric point for horseradish peroxidase isozymes ranges from 3.0 - 9.0.

Other Notes

View more information on peroxidase at www.sigma-aldrich.com/enzymeexplorer.

Packaging

5000, 25000, 50000, 100000, 200000 units in glass bottle

Unit Definition

One pyrogallol unit will form 1.0 mg purpurogallin from pyrogallol in 20 sec at pH 6.0 at 20 °C.

absorbance ratioRZ ≥1.0
formessentially salt-free, lyophilized powder
mol wt~44 kDa
Quality Level200
solubilityH2O: soluble, 0.1 M phosphate buffer: soluble (pH 6.0)
specific activity≥50 units/mg solid (using pyrogallol)
storage temp.2-8°C
typeType I
Cas Number9003-99-0
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