BrdU (5-Bromo-2′-deoxyuridine)

Thymidine analog which is incorporated into DNA during DNA replication (during S-phase of cell cycle). 

BrdU is used to identify proliferating cells. Labels cell lines and primary cell cultures in vitro and also cells in vivo. 

Widely used to study adult neurogenesis. Can be used in combination with neuron specific markers such as NeuN to identify newly formed neurons.

Frequently used to label and fate-map dividing cells in neural stem cell biology.

#Protocol 1: Serum starved vs stimulated BrdU proliferation assay

• HEK293T cells were cultured until approximately 50% confluency in 10% FBS in DMEM before media was exchanged to pure DMEM for 24hrs.
• Cells were either left in DMEM (serum starved) or incubated in 20% FBS in DMEM (serum starved + stimulated) for 2hrs in the prescence of 10µM BrdU (HB0978).
• Cells were fixed in 4% PFA and then stained for BrdU using a monoclonal anti-BrdU antibody (HB9919) following our ICC protocol then imaged using a confocal microscope.

#Protocol 2: Proliferation of HEK293T cells visualised using BrdU

• BrdU cells were cultured in 10% FBS in DMEM in the prescence of 10µM BrdU (HB0978) for 5 days.
• Cells were fixed in 4% PFA and then stained for BrdU using a monoclonal anti-BrdU antibody (HB9919) following our ICC protocol then imaged using a confocal microscope.