HRP
Horseradish Peroxidase
Cat. No.
BT13357
Source
Root extracts of horseradish.
Synonyms
Horseradish Peroxidase, HRP, EC 1.11.1.7.
Appearance
Sterile Filtered red-brown lyophilized powder.
Purity
(A403/A275) = RZ: 3.0.
Usage
Prospec's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
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Description
HRP consists of the basic isoenzyme having a molecular weight of 44 kDa.
The Horseradish Peroxidase is purified by affinity chromatography, which results in an enzyme of high specific activity and purity.
The Horseradish Peroxidase is purified by affinity chromatography, which results in an enzyme of high specific activity and purity.
Product Specs
Introduction
Horseradish peroxidase (HRP) is an enzyme extracted from horseradish that has significant applications in molecular biology, particularly in antibody amplification and detection. Its utility has made it a staple in neurobiology, surpassing even the Golgi stain in usage. HRP's versatility extends to techniques like Western blotting and ELISAs, where it serves as a crucial component. In immunoassays, HRP is frequently employed as an enzymatic label, often conjugated to antibodies, lectins, or haptens. This coupling process typically targets the carbohydrate side chains of the HRP molecule.
Description
HRP is primarily composed of the basic isoenzyme with a molecular weight of 44 kDa. Purification is achieved through affinity chromatography, yielding a highly specific and pure enzyme.
Physical Appearance
Red-brown lyophilized powder, sterile and filtered.
Solubility
For reconstitution of the lyophilized HRP, sterile 18MΩ-cm H2O is recommended, with a minimum concentration of 100 µg/ml.
Stability
While lyophilized HRP remains stable at room temperature for up to 3 weeks, storage in a desiccated state below -18°C is advised. After reconstitution, HRP should be stored at 4°C for a period of 2-7 days. For long-term storage, temperatures below -18°C are recommended. Repeated freeze-thaw cycles should be avoided.
Purity
The purity of HRP is represented by an A403/A275 ratio (RZ) of 3.0.
Biological Activity
The biological activity of HRP is determined to be 276 U/mg at 25°C and pH 7, using guaiacol as the hydrogen donor and H₂O₂ as substrates.
Synonyms
Horseradish Peroxidase, HRP, EC 1.11.1.7.
Source
Root extracts of horseradish.
Product Science Overview
Structure and Function
HRP is a metalloenzyme with multiple isoforms, the most studied being type C. It is a large alpha-helical glycoprotein that binds heme as a redox cofactor . The enzyme’s structure was first solved by X-ray crystallography in 1997 and has since been studied with various substrates . HRP catalyzes the conversion of chromogenic substrates (e.g., TMB, DAB, ABTS) into colored products and produces light when acting on chemiluminescent substrates (e.g., luminol) .
Applications
HRP is widely used in various biochemical assays due to its ability to amplify weak signals and increase the detectability of target molecules . Some common applications include:
- Western Blotting: HRP-conjugated antibodies are used to detect specific proteins. The enzyme produces a detectable signal in the presence of a substrate, allowing for the identification and quantification of proteins .
- ELISA (Enzyme-Linked Immunosorbent Assay): HRP is used to detect the presence of antigens or antibodies in a sample. The enzyme’s reaction with a substrate produces a color change, indicating the presence of the target molecule .
- Immunohistochemistry (IHC): HRP is used to visualize the presence and location of specific proteins in tissue sections. The enzyme’s reaction with a substrate produces a colored product that can be observed under a microscope .
Advantages
HRP is preferred in many applications due to its:
- High Turnover Rate: HRP has a high turnover rate, allowing for the generation of strong signals in a relatively short time span .
- Stability: HRP is more stable and less expensive compared to other enzymes like alkaline phosphatase .
- Versatility: HRP can be used with a variety of substrates, making it suitable for different types of assays .
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