How Do You Make Antibody Dilutions For IHC?
Immunohistochemistry (IHC) is a specialist technique used by scientists to visualize cells within tissues that have been placed on a slide and stained with an appropriate antibody that recognizes a protein that is unique to the cell or tissue under investigation. Antibody dilutions (titrations), must be performed using the experimental conditions specified in the user’s own protocol, not that of the antibody’s manufacturer, since the two may not be identical. Step 1 Check and prepare the antibody. Verify that the antibody has been raised in the correct host, and reacts with the exact version of the protein (known as the isoform, or splice variant) that is being tested. Using a pipette, carefully take a 10 microliter portion of this and label as ‘personal stock’. If only very small volumes of antibody are available, skip this step. Keeping a personal stock of antibody is routine practice in all laboratories as it prevents cross-contamination of the original stock. Dispense this into a l
