The optimal pH range for product P8375 is 6.0 - 6.5.
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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1000 UNITS
$70.20
2000 UNITS
$114.00
5000 UNITS
$130.00
10000 UNITS
$281.00
25000 UNITS
$480.00
100000 UNITS
$2,030.00
250000 UNITS
$3,890.00
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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
SMILES string
[O+H2]O[O-]
InChI
1S/H2O3/c1-3-2/h1-2H
InChI key
JSPLKZUTYZBBKA-UHFFFAOYSA-N
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1 of 4
This Item | P6782 | P2088 | P6140 |
|---|---|---|---|
| specific activity ≥250 units/mg solid (using pyrogallol) | specific activity ≥250 units/mg solid (using pyrogallol), 950-2000 units/mg solid (using ABTS) | specific activity 200-300 units/mg solid (using pyrogallol) | specific activity ≥225 units/mg protein (biuret, using pyrogallol) |
| application(s) diagnostic assay manufacturing | application(s) diagnostic assay manufacturing | application(s) - | application(s) - |
| form essentially salt-free, lyophilized powder | form essentially salt-free, lyophilized powder | form essentially salt-free, lyophilized powder | form ammonium sulfate suspension |
| solubility 0.1 M phosphate buffer: soluble 10 mg/mL, clear, orange to red (pH 6.0) | solubility 0.1 M phosphate buffer: soluble 10 mg/mL, clear, orange to red (pH 6.0), H2O: soluble | solubility - | solubility - |
| mol wt ~44 kDa | mol wt ~44 kDa | 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 |
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].
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.[4]
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.
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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signalword
Danger
hcodes
pcodes
Hazard Classifications
Resp. Sens. 1
Storage Class
11 - Combustible Solids
wgk_germany
WGK 1
ppe
Eyeshields, Gloves, type N95 (US)
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A Thermostable Hydrogen Peroxide Sensor Based on "Wiring" of Soybean Peroxidase
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The skeletal muscle T-tubule is a specialized membrane domain essential for coordinated muscle contraction. However, in the absence of genetically tractable systems the mechanisms involved in T-tubule formation are unknown. Here, we use the optically transparent and genetically tractable zebrafish
Articles
Discover our peroxidase from horseradish enzymes, products, substrates, and inhibitors for your ELISA, immunoassay, and protein application needs.
Protocols
This procedure is for the determination of Peroxidase enzymatic activity using Pyrogallol as the substrate.
To standardize a procedure for the assay of Peroxidase using 2,2'-Azino-bis(3-Ethylbenzthiazoline-6-Sulfonic Acid) as a substrate.
Reinheitszahl (RZ) is the ratio of absorbance due to hemin (A403, Soret region) to absorbance due to protein (A275).
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What is the optimal pH range of Peroxidase from horseradish, Product P8375?
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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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How do I convert units of Peroxidase from horseradish, Product P8375, to solid?
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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 can you tell me about the stability of Peroxidase from horseradish, Product P8375, (as is, in suspension and in solution)?
1 answer-
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.
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