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

Posted May 8, 2024


Antibodies and Proteomics
Bioconjugation
Antibody and Protein Labeling
Biotin and Streptavidin
Answer
  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

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