Thermo Fisher Scientific BrdU Monoclonal Antibody (BU20A), eBioscience

Thermo Fisher Scientific BrdU Monoclonal Antibody (BU20A), eBioscience

Applications and Tested Dilutions

Product Specifications

Additional Formats:

• APC
• eFluor 450
• FITC
• 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 the S-phase. Anti-BrdU antibodies are used to identify cells that have undergone DNA synthesis during BrdU treatment.

For flow cytometry staining, use the BrdU Staining Buffer Set (Product # 00-5525).

Applications Reported:

• Intracellular staining followed by flow cytometric analysis
• Immunohistology staining of frozen tissue sections

Applications Tested:

• Tested by flow cytometric analysis of BrdU-labeled mouse splenocytes
• Recommended ≤1 µg/test (100 µL final volume, 10⁵–10⁸ cells/test)

Recommended BrdU Labeling and Staining Protocol:

1. Label dividing cells with 10 µM BrdU for 45 min at 37°C.
2. Wash once with 1X PBS.
3. Stain surface molecules as per Surface Staining Protocol.
4. Wash in cold Flow Cytometry Staining Buffer or PBS.
5. Add 1 mL Foxp3 Fixation/Permeabilization solution (Product # 00-5521-00).
6. Incubate 30–60 min at 2–8°C in the dark.
7. Wash with 1X Permeabilization Buffer and centrifuge.
8. Resuspend in 100 µL Staining Buffer with 30 µg DNase I.
9. Incubate 1 hr at 37°C, wash.
10. Stain with anti-BrdU antibody 30–60 min, wash.
11. If necessary, stain with secondary antibody 30 min, wash.
12. Analyze by flow cytometry.

Purity: >90% (SDS-PAGE)
Aggregation: <10% (HPLC)
Filtration: 0.2 µm post-manufacturing filtered

Target Information

Bromodeoxyuridine (BrdU) is a synthetic nucleoside analog of thymidine used to detect proliferating cells. It incorporates into newly synthesized DNA during the S-phase, substituting thymidine. Detection requires DNA denaturation (acid or heat exposure) to allow antibody binding.

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