Peroxidase from horseradish, Type VI, essentially salt-free, lyophilized powder, 250-330 units/mg so

Peroxidase from horseradish, Type VI, essentially salt-free, lyophilized powder, 250-330 units/mg so

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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


Horseradish peroxidase (HRP) is isolated from horseradish roots (Amoracia rusticana) and belongs to the ferroprotoporphyrin group of peroxidases.. 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 P8375, type VI, is an essentially salt free lyophilized powder. It is commonly used to determine amounts of glucose and peroxides in solution. It has been used to study sensory input of cervical spinal cord neurons.


The enzyme has been used to develop a thermostable soybean peroxidase-based biosensor by cross-linking and electrically ?wiring? the enzyme through a redox-conducting hydrogel to a glassy carbon electrode. It has also been used to assay the oxidation of low density lipoprotein.


Biochem/physiol Actions


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. Known inhibitors are sodium azide, cyanide, L-cystine, dichromate, ethylenethiourea, hydroxylamine, sulfide, vanadate, p-aminobenzoic acid, and Cd2+, Co2+, Cu2+, Fe3+, Mn2+, Ni2+, and Pb2+ ions.


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 ?-galactosidase and alkaline phosphatase and hence, it is the most desired label. Also, its glycosylation leads to lower non-specific binding.


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


10000, 25000, 100000, 250000 units in glass bottle


1000, 2000 units in glass bottle

Quality Level300
ManufacturerSIGMA-ALDRICH
Storage Temp.2-8°C
Solubility0.1 M phosphate buffer: soluble 10 mg/mL, clear, orange to red (pH 6.0)
Formessentially salt-free, lyophilized powder

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