The methodology described in this course is from the Dako HercepTest™, as used with the Dako Classic instrumentation with antigen recovery being performed outside of the instrument. Antigen recovery is necessary to release the aldehyde bonds caused by formalin fixation. Heat-induced epitope retrieval (HIER) is performed in a water bath at 100º C for 40 minutes using a citrate buffer. Polymer technology is used to reduce biotin related background and to increase the signal. The polymer is based on dextran technology and consists of both secondary goat anti-rabbit IgG and horseradish peroxidase molecules linked to a common dextran polymer.
