AAT Bioquest

How does the streptavidin-biotin (LSAB) method work?

Posted May 8, 2024

  1. The tissue is exposed to the primary antibody, which attaches specifically to the antigen of interest.
  2. A biotin-labeled secondary antibody is introduced, binding to the primary antibody already attached to the antigen.
  3. An enzyme-labeled streptavidin complex is applied, where streptavidin directly binds to enzyme molecules. This complex then binds to the biotin-labeled secondary antibody.
  4. Lastly, a chromogenic substrate is added to reveal the peroxidase enzyme, allowing visualization of the antigen site.
Additional resources

Immunohistochemistry in investigative and toxicologic pathology

Biotin and Streptavidin

HRP-Biotin Conjugate

Streptavidin Concentration Calculator