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Cat. No.: HPA-0.2 (for 0.2ml)

Cat. No.: HPA-1 (for 1ml)

Cat. No.: HPAL-1 (for 1mg)

Cat. No.: HPAL-5 (for 5mg)

Description

Horseradish Peroxidase-labeled Protein A (also referred to as HRP-labeled Protein A, HRP-Protein A, Protein A-HRP, Protein A (HRP), or Protein A (HRP Conjugate)) is designed for the detection of IgG and IgG-conjugated or IgG-bound antibodies, nucleic acids, proteins, and other biomolecules. It is widely applicable in techniques such as Western blotting, ELISA, and immunohistochemistry (IHC).

This HRP-Protein A conjugate is synthesized by covalently linking highly purified recombinant Protein A (>98% purity by SDS-PAGE and HPLC) with ultrapure horseradish peroxidase (RZ >3), ensuring minimal background and exceptional sensitivity.

Protein A is a cell wall component derived from several strains of Staphylococcus aureus. It is a single polypeptide chain with minimal carbohydrate content. Recombinant Protein A, produced in E. coli, retains the functional properties of native Protein A. It consists of five IgG-binding domains (E-D-A-B-C) arranged in tandem, with the cell wall binding region, membrane binding region, and albumin binding region removed to maximize specificity for IgG. Protein A binds strongly to various subclasses of human, mouse, and rat IgG (e.g., human IgG1, IgG2, IgG4; mouse IgG2a, IgG2b, IgG3; rat IgG2c), and also to total IgG from rabbit, guinea pig, pig, dog, and cat. This makes it an ideal tool for purifying both polyclonal and monoclonal IgG antibodies.

Horseradish Peroxidase (HRP) catalyzes chemiluminescent substrates (e.g., ECL, BeyoECL) in Western blotting, EMSA, Southern, or Northern blotting. In ELISA, it  catalyzes TMB to produce a blue color, and in IHC, immunocytochemistry, or Western blotting, it catalyzes DAB to yield a brown precipitate.

Application

HRP-Protein A is available in two formats: liquid and lyophilized powder.

• Liquid format: Ready-to-use at 1 mg/mL.

• Lyophilized powder: Reconstitute with ultrapure water or PBS (pH 7.4) to a recommended concentration of 1 mg/mL.

The following table provides recommended dilution ranges for common applications. Optimal dilution should be determined based on specific experimental conditions, including detection sensitivity, antibody affinity, incubation temperature, and duration.

Storage

• Store at –20°C, protected from light. Stable for 1 year under these conditions.
• Lyophilized powder: Store at 4°C, stable for at least 3 months.
• Liquid formulation: Store at 4°C, stable for at least 1 week.

Precautions

• Upon receipt, use the product immediately or store it under the recommended conditions. Enzymatic activity may gradually decline over time.
• To avoid loss of activity due to repeated freeze-thaw cycles, it is recommended to aliquot the solution after initial reconstitution and store at –20°C.
• Sodium azide inhibits horseradish peroxidase activity and should not be added to solutions containing HRP.
• When reconstituting the lyophilized powder, gently pipette up and down or swirl to fully dissolve.
• The lyophilized powder contains protective stabilizers; the 1 mg/mL liquid formulation contains 50% glycerol.
• Do not vortex vigorously, as this may denature the protein and reduce activity.
• For research use only. Not for clinical diagnosis or therapeutic use. Not for use in food, drugs, or household applications.
• For your safety, wear a lab coat and disposable gloves when handling this product.

Related:

• Horseradish Peroxidase-labeled Protein A (HRP-Protein A)
• Horseradish Peroxidase-labeled Protein G (HRP-Protein G)
• Horseradish Peroxidase-labeled Protein A/G (HRP-Protein A/G)
• Horseradish Peroxidase-labeled Protein L (HRP-Protein L)
   

Only for research and not intended for treatment of humans or animals

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