HRP Antibodies
Horseradish Peroxidase (HRP) antibodies are conjugated antibodies that are chemically linked to horseradish peroxidase, an enzyme widely used in biochemical assays to catalyze the oxidation of substrates, producing a colorimetric, chemiluminescent, or fluorescent signal. HRP is frequently used as a labeling agent in secondary antibodies due to its stability, cost-effectiveness, and ability to generate strong signals.
Content of HRP Antibodies
HRP antibodies are typically secondary antibodies conjugated with the HRP enzyme. These antibodies are produced by immunizing a host species (commonly rabbit, goat, or mouse) with an antigen from another species, followed by chemical conjugation of the antibody with the HRP enzyme.
Key specifications:
- Immunogen: Antigens from species-specific immunoglobulin (e.g., anti-mouse IgG, anti-rabbit IgG).
- Host: Typically raised in rabbits, goats, or mice.
- Conjugation: Covalently linked to horseradish peroxidase (HRP) enzyme.
- Applications: Suitable for Western blotting (WB), enzyme-linked immunosorbent assay (ELISA), immunohistochemistry (IHC), immunocytochemistry (ICC), and chemiluminescent assays.
- Reactivity: Typically recognizes immunoglobulins from species such as human, mouse, rabbit, rat, etc.
Application of HRP Antibodies
HRP-conjugated antibodies are extensively used in immunoassays due to the enzyme's ability to produce a detectable signal when a substrate such as 3,3′,5,5′-tetramethylbenzidine (TMB) or enhanced chemiluminescent (ECL) reagents are added. The applications include:
- Enzyme-Linked Immunosorbent Assay (ELISA): HRP antibodies are critical in ELISA, where they bind to the primary antibody or antigen on a solid surface. The addition of a substrate, such as TMB, results in a colorimetric change that quantifies the presence of the antigen or antibody being detected.
- Immunohistochemistry (IHC): HRP-conjugated antibodies are used in IHC to detect the presence of specific proteins in tissue sections. HRP catalyzes a chromogenic substrate reaction, creating a localized color change that reveals the tissue distribution of the target antigen.
- Immunocytochemistry (ICC): HRP antibodies can be used in ICC for detecting antigens within cells, allowing researchers to visualize protein localization using light microscopy after a substrate reaction has occurred.
The use of HRP-conjugated antibodies significantly enhances assay sensitivity and visualization, making them indispensable in molecular biology, diagnostics, and research. HRP antibodies provide robust detection and amplification of the target antigen, enabling researchers to achieve accurate results in various immunoassay platforms.
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