BrdU incorporation for raft cultures

Contributed by Matt Lewis <hop2kin@yahoo.co.uk>

BrdU incorporation for raft cultures

• Prepare stocks of BrdU at 10mg/ml in H2O, filter sterilise and store in working aliquots (eg. 200 microlitres) at –20C.
• Incubate rafts with BrdU (100 microgram per ml) for 12 hours prior to harvesting.
• Detect incorporated BrdU via antibody staining (Zymed 18-0103) using standard immunohistochemical techniques

High temperature antigen retrieval is required to open up the DNA

This method is based, with permission, on an original protocol available here.