The protocol involves labeling cells with brdu, followed by fixation, dna denaturation (to expose brdu epitopes), and immunostaining. Nalog that gets incorporated into a cell's dna when the cell is dividing Brdu is commonly used in the det Ction of proliferating cells in living tissues or as a cell cycle marker Brdu is characterized as a mutagenic. Immunohistochemistry (ihc) is currently one of the most popular histological techniques, which is widely used to recognize and detect antigens in different tissues
The principle behind the ihc is the specific binding of antibodies to target antigenic epitopes and their subsequent detection. In order for brdu to be accessible for the antibody to bind, the dna has to be denatured If a cell is positive for brdu, it was going through s phase when brdu was present Normally, dna has to be denatured for the brdu antibody to detect any brdu incorporated into the dna. In the brdu assay, cells are incubated with brdu (synthetic analog of thymidine, incorporated in cellular dna during replication) Brdu antibodies can be used in conjunction with cell type markers such as ki67, doublecortin, and neun to identify proliferating cells and newly differentiated neurons.
