BrdU Monoclonal Antibody (BU20A), eBioscience™, Invitrogen™

Description

• Description: This Bu20a monoclonal antibody reacts with 5-bromodeoxyuridine (BrdU)
• BrdU is a derivative of uridine that can be incorporated into DNA in place of thymidine during the S-phase of the cell cycle
• Anti-BrdU can then be used to identify cells that have undergone DNA synthesis during BrdU treatment
• For staining for flow cytometric analysis, we recommend the use of the BrdU Staining Buffer Set (cat
• 00-5525) and protocol
• Applications Reported: This Bu20A antibody has been reported for use in intracellular staining followed by flow cytometric analysis and immunohistology staining of frozen tissue sections
• Applications Tested: This Bu20A antibody has been tested by flow cytometric analysis of BrdU-labeled mouse splenocytes
• This can be used at less than or equal to 1 μg per test
• A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL
• Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test
• It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest
• BrdU labeling and staining with anti-BrdU antibody:1
• Label dividing cells with 10 μM BrdU for 45 min at 37°C.2
• Following the incubation, harvest the cells and wash once with 1X PBS.3
• Stain surface molecules according to the Surface Staining Protocol.4
• Wash in cold Flow Cytometry Staining Buffer or 1X PBS.5
• Bromodeoxyuridine (5-bromo-2-deoxyuridine, BrdU) is a synthetic nucleoside that is an analogue of thymidine
• BrdU is commonly used in the detection of proliferating cells in living tissues, and can be incorporated into the newly synthesized DNA of replicating cells (during the S phase of the cell cycle), substituting for thymidine during DNA replication
• Antibodies specific for BrdU can then be used to detect the incorporated chemical, thus indicating cells that were actively replicating their DNA
• Binding of the antibody requires denaturation of the DNA, usually by exposing the cells to acid or heat.