Immediately label your main antibody with HRP

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

The enzyme label may very well be visualized through chromogenic reactions. For example, diaminobenzidine (DAB) inside the presence of hydrogen peroxide (H₂0₂) is reworked proper right into a water-insoluble brown pigment. Completely different substrates that may be utilized to measure horseradish peroxidase train embrace ABTS, TMB, and TMBUS.

HRP is a popular detection label utilized in evaluation. Antibody-HRP conjugates are typically utilized in ELISA, IHC, and western blotting. HRP may very well be conjugated to the primary antibody for direct detection or secondary antibody for indirect detection.

Direct detection is normally a preferred methodology inside these functions to avoid cross-species reactivity and to eradicate additional laborious wash and separation steps, notably in time-consuming protocols. Nonetheless, immediately conjugating HRP to the antibody or protein of choice may very well be powerful and labor-intensive if using typical methodology.