Direct detection is often a preferred method within these applications to avoid cross-species reactivity and to eliminate additional laborious wash and separation steps, especially in time-consuming protocols. However, directly conjugating HRP to the antibody or protein of choice can be difficult and labor-intensive if using traditional methodology.

Our Lightning-Link HRP antibody labeling kit allows the direct conjugation of your antibody, protein, or peptide to HRP for use in any application. The kit only requires 30 seconds of hands-on time, and conjugates are ready to use after only 3 hours. Furthermore, there is no need for extra wash or separation steps post conjugation.

​Directly label your primary antibody with HRP

​​​Horseradish peroxidase (HRP) is a 44 kDa glycoprotein with 6 lysine residues, which can be conjugated to antibodies and proteins for use in a variety of applications.

The enzyme label can be visualized through chromogenic reactions. For example, diaminobenzidine (DAB) in the presence of hydrogen peroxide (H₂0₂) is converted into a water-insoluble brown pigment. Other substrates that can be used to measure horseradish peroxidase activity include ABTS, TMB, and TMBUS.

HRP is a popular detection label used in research. Antibody-HRP conjugates are commonly used in ELISA, IHC, and western blotting. HRP can be conjugated to the primary antibody for direct detection or secondary antibody for indirect detection.

Benefits of using Lightning-Link HRP

• Quick and easy – 30 seconds of hands-on time
• Adjustable ratios of antibody to HRP, between 1:1 and 1:4
• No separation steps involved, recover 100% of your antibody or protein
• Potentially more sensitive when compared to traditional methods – so less antibody required (see figure 1 below)
• Label from as little as 10 µg – scale up to a gram or more.

Abcam’s HRP secondaries advantages are:

• High sensitivity – detect low-abundant targets with high signal-to-noise ratios due to an optimal number of HRP molecules per antibody.
• Robustness – can be used at dilutions ranging from 1/2,000 to 1/20,000 and provide high specificity and minimal background.
• Stability – they are supplied in a liquid format that is ready-to-use and can be stored at 4°C.
• Flexibility – compatible with chromogenic, fluorogenic and chemiluminescent substrates.
• Cost-effective – use less conjugate per assay and perform more blots.