Invitrogen™ BrdU Monoclonal Antibody (BU20A), APC, 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. Applications Tested: This BU20A antibody has been pre-titrated and tested by intracellular staining and flow cytometric analysis of BrdU-labeled mouse splenocytes using the Foxp3/Transcription Factor Buffer Set (cat. 00-5523) and protocol or the BrdU Staining Buffer Set (cat. 00-5525) and protocol. Please see Best Protocols Section (Staining intracellular Antigens for Flow Cytometry) for staining protocol (refer to Protocol B: One-step protocol for intracellular (nuclear) proteins). This can be used at 5 μL (0.125 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. BrdU labeling and staining with the Anti-BrdU antibody:1. Label dividing cells with 10 μM BrdU for 45 min at 37°C.2.

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.