How to Perform Immunohistochemistry Using Biotinylated Peptide?

A peptide RK-10 is used to detect Programmed Death Ligand 1 (PD-L1) expressing tumors with immunohistochemistry.

  1. In order to detect PD-L1 in tissues, a biotin-conjugated version of peptide RK-10-Cy5 was used.
  2. Tissues were incubated with 15 µM biotinylated peptide for 2 hours.
  3. After 2 hours, slides were washed with buffer and treated with Pierce™ High Sensitivity Streptavidin-HRP (1:200 dilution) for 30 minutes.
  4. Once this was complete, slides were again washed in buffer then treated with DAB (Sigma) for 10 minutes. Slides were again washed in buffer, then dehydrated using graded alcohol and xylene and counterstained with hematoxylin.
  5. Slides were then imaged using bright-field microscopy on a Leica DM5500.