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P8375

Peroxidase from horseradish

Type VI, essentially salt-free, lyophilized powder, ≥250 units/mg solid (using pyrogallol)

Synonym(s):

Detection Enzyme, HRP, Peroxidase, Donor:hydrogen-peroxide oxidoreductase, Horseradish peroxidase

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About This Item

CAS Number:
9003-99-0
eCl@ss:
32160410
UNSPSC Code:
12352204
NACRES:
NA.54
EC Number:
232-668-6
MDL number:
MFCD00071339
EC Number:
1.11.1.7 (BRENDA, IUBMB)
Specific activity:
≥250 units/mg solid (using pyrogallol)

Key Documents

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Product Name

Peroxidase from horseradish, Type VI, essentially salt-free, lyophilized powder, ≥250 units/mg solid (using pyrogallol)

SMILES string

[O+H2]O[O-]

InChI key

JSPLKZUTYZBBKA-UHFFFAOYSA-N

InChI

1S/H2O3/c1-3-2/h1-2H

type

Type VI

form

essentially salt-free, lyophilized powder

specific activity

≥250 units/mg solid (using pyrogallol)

mol wt

~44 kDa

solubility

0.1 M phosphate buffer: soluble 10 mg/mL, clear, orange to red (pH 6.0)

absorbance ratio

RZ 2.5-4.0

application(s)

diagnostic assay manufacturing

storage temp.

2-8°C

Quality Level

300

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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

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

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

One pyrogallol unit will form 1.0 mg purpurogallin from pyrogallol in 20 sec at pH 6.0 at 20 °C.
View more information on peroxidase at www.sigma-aldrich.com/enzymeexplorer.

pictograms

Health hazard
GHS08

signalword

Danger

hcodes

H334

Hazard Classifications

Resp. Sens. 1

Storage Class

11 - Combustible Solids

wgk

WGK 1

ppe

Eyeshields, Gloves, type N95 (US)

Regulatory Listings

Regulatory Listings are mainly provided for chemical products. Only limited information can be provided here for non-chemical products. No entry means none of the components are listed. It is the user’s obligation to ensure the safe and legal use of the product.

P8375-10MG: + P8375-BULK: + P8375-25KU: + P8375-5KU: + P8375-2KU: + P8375-250KU: + P8375-100KU: + P8375-1KU: + P8375-10KU: + P8375-VAR:

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Certificates of Analysis (COA)

Lot/Batch Number
0000488641
0000488639
0000488640
0000488639
0000488640

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A Thermostable Hydrogen Peroxide Sensor Based on "Wiring" of Soybean Peroxidase
Analytical Chemistry, 67 (23), 4247-4249 (1995)
Filip Mollerup et al.
PloS one, 14(5), e0216546-e0216546 (2019-05-16)
Copper radical alcohol oxidases belonging to auxiliary activity family 5, subfamily 2 (AA5_2) catalyze the oxidation of galactose and galactosides, as well as aliphatic alcohols. Despite their broad applied potential, so far very few AA5_2 members have been biochemically characterized.
Krishna Mohan Pathi et al.
Frontiers in plant science, 11, 543895-543895 (2020-11-17)
Biotic stresses caused by microbial pathogens impair crop yield and quality if not restricted by expensive and often ecologically problematic pesticides. For a sustainable agriculture of tomorrow, breeding or engineering of pathogen-resistant crop varieties is therefore a major cornerstone. Maize
Marilyn S Lee et al.
Synthetic and systems biotechnology, 5(3), 145-154 (2020-07-09)
Cell-free systems contain many proteins and metabolites required for complex functions such as transcription and translation or multi-step metabolic conversions. Research into expanding the delivery of these systems by drying or by embedding into other materials is enabling new applications
E Wieland et al.
Proceedings of the National Academy of Sciences of the United States of America, 90(13), 5929-5933 (1993-07-01)
Oxidative modification of low density lipoprotein is believed to be an important pathway by which the lipoprotein becomes atherogenic. The in vitro systems for oxidative modification of low density lipoprotein thus far described all appear to depend upon the presence
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Articles

Horseradish Peroxidase (HRP) Enzymes

Discover our peroxidase from horseradish enzymes, products, substrates, and inhibitors for your ELISA, immunoassay, and protein application needs.

Protocols

ペルオキシダーゼ（EC 1.11.1.7）のイムノアッセイ：基質としての2,2'-アジノ-ビス（3-エチルベンズチアゾリン-6-スルホン酸）

2,2'-アジノ-ビス（3-エチルベンズチアゾリン-6-スルホン酸）を基質として用いたペルオキシダーゼのアッセイ手順の標準化

Enzymatic Assay of Peroxidase (EC 1.11.1.7)

This procedure is for the determination of Peroxidase enzymatic activity using Pyrogallol as the substrate.

Enzymatic Assay of Peroxidase (EC 1.11.1.7) 2,2'-Azino-bis(3-Ethylbenzthiazoline-6-Sulfonic Acid) as a Substrate

To standardize a procedure for the assay of Peroxidase using 2,2'-Azino-bis(3-Ethylbenzthiazoline-6-Sulfonic Acid) as a substrate.

Spectrophotometric Determination of Reinheitszahl (RZ) for Peroxidase (EC 1.11.1.7)

Reinheitszahl (RZ) is the ratio of absorbance due to hemin (A403, Soret region) to absorbance due to protein (A275).

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Data Sheet - P8375

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