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HRP Conjugated Antibodies & Reagents

Horseradish peroxidase (HRP) is a metalloenzyme found in the root of the horseradish plant, where it supports essential processes such as lignification and pathogen defense by using hydrogen peroxide (H2O2) to oxidize a diverse range of compounds.

HRP is widely used as a reporter enzyme for scientific research due to its high turnover rate, which enables the rapid generation of strong signals that can enhance the detection of low abundance targets. Other advantages of HRP include its relatively small size (~44 kDa), which facilitates sample penetration, and its stability towards external factors such as temperature and organic solvents.

Applications in which HRP is used include enzyme-linked immunosorbent assay (ELISA), ELISpot, and Western blotting, as well as immunohistochemistry (IHC) and immunocytochemistry (ICC). These typically involve using HRP-conjugated antibodies to detect a target of interest, followed by the addition of an appropriate substrate to generate a measurable signal.

We offer an extensive selection of HRP-conjugated primary and secondary antibodies, including F(ab)2 and Fab fragments. In addition, our product portfolio encompasses HRP-conjugated isotype controls, Protein A/G-HRP, and HRP-streptavidin conjugates.

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Frequently Asked Questions

HRP-conjugated antibodies are produced by covalently attaching multiple HRP molecules to primary or secondary antibodies. Because the HRP enzyme activity is maintained, it can be used for confirming whether the antibody has detected its antigenic target. This is accomplished by adding a peroxidase substrate at the end of the immunostaining workflow, since HRP does not produce a measurable signal. Common colorimetric substrates include TMB and ABTS, which are commonly used for ELISA, and DAB, which is widely used for IHC. For fluorometric detection, ADHP is popular. In addition, many different chemifluorescent and chemiluminescent substrates are commercially available.

HRP-conjugated antibodies are used in many different research applications. These include ELISA, ELISpot, Western blotting, IHC, and ICC, all of which require careful optimization of the staining protocol to ensure accurate results.

Factors to consider when selecting an HRP-conjugated antibody will vary depending on whether you wish to perform direct or indirect detection. For direct detection (using an HRP-conjugated primary antibody), the antibody should be validated for your chosen species and application. You may also wish to decide whether to use a polyclonal antibody, which can provide signal amplification, or a monoclonal antibody, which offers the advantage of unlimited supply. The antibody host species is another key factor, especially if you intend to analyze multiple targets in the same experiment, using a combination of direct and indirect detection.

For indirect detection (using an unlabeled primary antibody and an HRP-conjugated secondary antibody), it is critically important to select a secondary antibody that recognizes the host species of the primary antibody. Application-specific validation data is desirable but not essential, since secondary antibodies tend to work in most experimental settings. However, you should always perform testing in your own model system prior to using the antibody with precious samples.

HRP is purified from horseradish root, meaning that each preparation will have a slightly different activity level depending on environmental conditions. To account for this, we dilute our HRP-conjugated antibodies and reagents to potency following quality testing to help ensure consistency between lots when using a similar working dilution. As a result, each HRP-conjugated product has a lot-specific concentration.

All SouthernBiotech HRP-conjugated antibodies and reagents are quality tested in-house by ELISA to ensure target specificity and lot-to-lot consistency.

HRP-conjugated antibodies and reagents can be stored at either 2-8°C or -20°C. There is no need to aliquot HRP-conjugated antibodies and reagents for storage at -20°C, since the buffer in which they are supplied contains 50% glycerol. Storage at -80C should be avoided as this will incur a freeze/thaw cycle that could compromise HRP enzyme activity.

Related Product Categories

Conjugated Antibodies

For the direct detection of your target, explore our offering of over 25 enzyme and fluorochrome conjugated primary antibodies.

Immunohistochemistry Antibodies

Enhance your research with our in-house produced and validated antibodies for IHC assays.

Primary Antibodies

Find a high-quality primary antibody from our fast-shipping selection, all produced to our exacting standards in our own facility.

Secondary Antibodies & Reagents

With a wide range of ready-to-ship secondary antibodies that are second to none, SouthernBiotech will keep your assays running and projects on schedule.