Peroxidase from horseradish
SIGMA/P8250 - Type II, essentially salt-free, lyophilized powder, 150-250 units/mg solid (using pyrogallol)
Synonym: Donor:hydrogen-
CAS Number: 9003-99-0
EC Number: 232-668-6
MDL Number: MFCD00071339
Product Type: Chemical
This picture is provided solely for illustration purposes. Optical properties of the actual product may deviate. Relevant product information is printed on labeled products and other accompanying or available information material. This image depicts SKU: P8250-25KU
This picture is provided solely for illustration purposes. Optical properties of the actual product may deviate. Relevant product information is printed on labeled products and other accompanying or available information material. This image depicts SKU: P8250-100KU
This picture is provided solely for illustration purposes. Optical properties of the actual product may deviate. Relevant product information is printed on labeled products and other accompanying or available information material. This image depicts SKU: P8250-50KU
This picture is provided solely for illustration purposes. Optical properties of the actual product may deviate. Relevant product information is printed on labeled products and other accompanying or available information material. This image depicts SKU: P8250-200KU
This picture is provided solely for illustration purposes. Optical properties of the actual product may deviate. Relevant product information is printed on labeled products and other accompanying or available information material. This image depicts SKU: P8250-5KU
This picture is provided solely for illustration purposes. Optical properties of the actual product may deviate. Relevant product information is printed on labeled products and other accompanying or available information material. This image depicts SKU: P8250-500KU
| absorbance ratio | RZ ≥1.8 |
| application(s) | diagnostic assay manufacturing |
| biological source | horseradish |
| form | essentially salt-free, lyophilized powder |
| InChI | 1S/H2O3/c1-3-2/h1-2H |
| InChI key | JSPLKZUTYZBBKA-UHFFFAOYSA |
| mol wt | ~44 kDa |
| Quality Level | 300  |
| SMILES string | [O+H2]O[O-] |
| solubility | 0.1 M phosphate buffer: soluble (pH 6.0) |
| H2O: soluble | |
| specific activity | 150-250 units/mg solid (using pyrogallol) |
| storage temp. | 2-8°C |
| type | Type II |
| Analysis Note: | Preliminary work shows it to contain at least five isoenzymes. |
| 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). 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. Horseradish peroxidase, product P8250, has been used to study nonoral antigens in inflamed gingiva and Ebola virus glycoprotein toxicity. |
| Application: | The enzyme has been used as a comparison during the peroxidase assay of extract from mature tall fescue leaf blades. It has also been used to measure H2O2 production. |
| 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 found to inhibit the enzyme activity. |
| 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. |
| 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. |
| Other Notes: | View more information on peroxidase at www.sigma-aldrich.com/enz. |
| Packaging: | 5000, 25000, 50000, 100000, 200000 units in glass bottle |
| Symbol |  GHS08 |
| Signal word | Danger |
| Hazard statements | H334 |
| Precautionary statements | P261 - P284 - P304 + P340 + P312 - P501 |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 3 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| activity | specific activity: 150-250 units/mg solid (using pyrogallol) |
| Storage Temp. | 2-8°C |
| Enzyme Commission (EC) Number | 1.11.1.7 ( BRENDA  | IUBMB ) |
| UNSPSC | 12352204 |