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Can I use an HRP-conjugated primary antibody for western blot (with no secondary)?
Posted April 24, 2020
Antibodies and Proteomics Bioconjugation Chemical ReagentsAntibody and Protein LabelingHRPHRP-labeled primary antibodyHorseradish Peroxidase (HRP) and Poly-HRPPrimary AntibodiesSecondary AntibodiesWestern Blotantibody conjugation
Answer
Yes, it is possible to use an HRP-conjugated primary antibody to perform a direct western blot, instead of performing an indirect assay with a conjugated secondary antibody. However, this should only be done if signal amplification is not needed, or else the signal may be too low. In this case, using a conjugated secondary antibody instead will help to improve the signal.
Additional resources
HRP-labeled goat anti-rabbit IgG (H+L)Bass, J. J., Wilkinson, D. J., Rankin, D., Phillips, B. E., Szewczyk, N. J., Smith, K., & Atherton, P. J. (2017). An overview of technical considerations for Western blotting applications to physiological research. Scandinavian journal of medicine & science in sports, 27(1), 4-25. doi:10.1111/sms.12702Western Blotting Assays