This product is stable as a powder, kept refrigerated. See the certificate of analysis for a lot-specific recommended retest date. It is also stable as a crystalline suspension of 3.2 M (NH4)2SO4 solution containing potassium phosphate buffer, pH 6.0 (see Product No. P6140). It is reasonably stable in solution in phosphate buffer. We have found that at 10 mg/mL in 0.1 phosphate buffer, pH 6.0, solutions kept frozen at -20°C lose less than 2% of their activity per week.
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:
eCl@ss:
32160410
UNSPSC Code:
12352204
NACRES:
NA.54
EC Number:
232-668-6
MDL number:
Specific activity:
≥250 units/mg solid (using pyrogallol)
Pricing and availability is not currently available.
type
Type VI
Quality Segment
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
SMILES string
[O+H2]O[O-]
InChI
1S/H2O3/c1-3-2/h1-2H
InChI key
JSPLKZUTYZBBKA-UHFFFAOYSA-N
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.
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[1].
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.[2]
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 β-galactosidase and alkaline phosphatase and hence, it is the most desired label. Also, its glycosylation leads to lower non-specific binding.
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.
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.
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This Item | |||
|---|---|---|---|
| specific activity ≥250 units/mg solid (using pyrogallol) | specific activity ≥250 units/mg solid (using pyrogallol) | specific activity ≥50 units/mg solid (using pyrogallol) | specific activity 200-300 units/mg solid (using pyrogallol) |
| form essentially salt-free, lyophilized powder | form powder, stabilized | form essentially salt-free, lyophilized powder | form essentially salt-free, lyophilized powder |
| application(s) diagnostic assay manufacturing | application(s) diagnostic assay manufacturing | application(s) - | application(s) - |
| mol wt ~44 kDa | mol wt - | mol wt ~44 kDa | mol wt ~44 kDa |
| storage temp. 2-8°C | storage temp. 2-8°C | storage temp. 2-8°C | storage temp. 2-8°C |
| solubility 0.1 M phosphate buffer: soluble 10 mg/mL, clear, orange to red (pH 6.0) | solubility phosphate buffer: soluble | solubility 0.1 M phosphate buffer: soluble (pH 6.0), H2O: soluble | solubility - |
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signalword
Danger
hcodes
pcodes
Hazard Classifications
Resp. Sens. 1
Storage Class
11 - Combustible Solids
wgk
WGK 1
ppe
Eyeshields, Gloves, type N95 (US)
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What can you tell me about the stability of Peroxidase from horseradish, Product P8375, (as is, in suspension and in solution)?
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How do I convert units of Peroxidase from horseradish, Product P8375, to solid?
1 answer-
The activity listed on the Certificate of Analysis for each lot can be used to convert from units to milligrams of solid.
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What is the Department of Transportation shipping information for this product?
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Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
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What is the optimal pH range of Peroxidase from horseradish, Product P8375?
1 answer-
The optimal pH range for product P8375 is 6.0 - 6.5.
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