Thermo Fisher Scientific BrdU Monoclonal Antibody (BU20A), FITC, eBioscience

Thermo Fisher Scientific BrdU Monoclonal Antibody (BU20A), FITC, eBioscience

Applications and Tested Dilution

Product Specifications

Additional Formats

• Unconjugated
• APC
• eFluor 450
• PE
• PerCP-eFluor 710
• Request custom conjugation

Product Specific Information

This BU20A monoclonal antibody reacts with 5-bromodeoxyuridine (BrdU), a uridine derivative incorporated into DNA during S-phase. Anti-BrdU antibodies identify cells undergoing DNA synthesis during BrdU treatment.

For flow cytometry staining, use BrdU Staining Buffer Set (Product #00-5525) and protocol.
This antibody is validated for intracellular staining in flow cytometry and immunohistology of frozen tissue sections.

Recommended Usage

• Tested by intracellular staining and flow cytometric analysis of BrdU-labeled mouse splenocytes using Foxp3/Transcription Buffers (Product #00-5521-00) or BrdU Staining Buffer Set (Product #00-5525-00).
• Refer to BestProtocols® for intracellular antigen staining (Protocol B: One-step protocol for nuclear proteins).
• Use at 5 µL (1 µg) per test; one test defined as staining a cell sample in 100 µL final volume (10⁵–10⁸ cells/test).
• Antibody titration recommended for optimal performance.

BrdU Labeling and Staining Protocol (Flow Cytometry)

1. Label dividing cells with 10 µM BrdU for 45 min at 37°C.
2. Harvest and wash cells with 1X PBS.
3. Perform surface staining as per protocol.
4. Wash in cold Flow Cytometry Staining Buffer or PBS.
5. Resuspend pellet and add 1 mL Foxp3 Fixation/Permeabilization Buffer (Product #00-5523-00).
6. Incubate 30–60 min at 2–8°C in the dark.
7. Wash and centrifuge; decant supernatant.
8. Resuspend pellet in 100 µL Flow Cytometry Staining Buffer with 30 µg Dnase I.
9. Incubate 1 hr at 37°C, then wash.
10. Stain with anti-BrdU antibody for 30–60 min, wash, and analyze.

Excitation: 488 nm
Emission: 520 nm
Laser: Blue Laser
Filtration: 0.2 µm post-manufacturing filtered

Target Information

Bromodeoxyuridine (5-bromo-2-deoxyuridine, BrdU) is a synthetic nucleoside and thymidine analogue. It is incorporated into newly synthesized DNA of replicating cells during S-phase, enabling detection of proliferating cells. Anti-BrdU antibody binding requires DNA denaturation via acid or heat exposure.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without authorization.

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