HRP-conjugated IHC secondary antibodies
Our HRP-conjugated IHC secondary antibodies are designed to provide robust and reliable signaling in immunohistochemistry (IHC) applications. These antibodies are conjugated to horseradish peroxidase (HRP), enabling highly sensitive detection through enzymatic color development. They offer specific binding to primary antibodies, amplifying the signal to facilitate the visualization of target antigens in tissue sections.
Our extensive portfolio of HRP-conjugated secondary antibodies offers many options for each target species. Search by popular target species to find the antibody that meets your needs.
Immunohistochemistry (IHC) is a widely used technique for the visualization of the presence and localization of specific proteins in tissue sections.
We offer a wide range of secondary antibodies validated for IHC that are available against several species and come conjugated with various reporter molecules (eg HRP) for easy colorimetric detection. Our secondary antibodies are suitable for use on various tissue samples, such as frozen and paraffin-embedded tissues.
Enzymatic detection methods
Enzymatic detection methods use horseradish peroxidase (HRP) or alkaline phosphatase (AP) conjugated secondary antibodies allowing colorimetric with different substrates for a range of applications.
Chromogenic detection in IHC
The enzymes used to detect antigens in chromogenic detection are typically conjugated to secondary antibodies, although primary antibodies directly conjugated to the enzyme can also be used. The most commonly used enzymes are horseradish peroxidase (HRP), which converts 3,3’-diaminobenzidine (DAB) into a brown product, and alkaline phosphatase (AP), which converts 3-amino-9-ethyl carbazole (AEC) into a red product. However, the choice of chromogen is determined by the enzyme used.
Chromogenic detection is usually more sensitive than fluorescent detection due to the higher signal amplification. Furthermore, unlike fluorophores, the colored precipitates created from substrates such as DAB are photostable, enabling storage of the slides for many years.
HRP-conjugated IHC secondary antibodies
Our HRP conjugated secondary antibodies are available for diverse species and isotypes (eg Goat anti-Mouse IgG, Goat anti-Rabbit IgG, Rabbit anti-Human IgG). We also have a range of HRP conjugated secondaries in fragment forms (e.g., F(ab')2) for applications requiring reduced Fc-mediated interactions. In addition, we offer pre-adsorbed options that reduce cross-species reactivity to minimize non-specific background in IHC staining.
For each application, HRP-conjugated secondary antibodies need substrates that can deliver high signal-to-noise ratios for the chosen detection method.
Pre-adsorbed HRP-conjugated secondary antibodies
Pre-adsorption (also referred to as cross-adsorption) is an extra purification step introduced to increase the specificity of a secondary antibody. The solution containing secondary antibodies is passed through a column matrix containing immobilized serum proteins from potentially cross-reactive species. Non-specific secondary antibodies are retained in the column, while highly specific secondaries flow through.
We recommend using pre-adsorbed (cross-absorbed) HRP- HRP-conjugated secondary antibodies to minimize non-specific background when working with tissues and cells.