Thermo Fisher Scientific BrdU Monoclonal Antibody (BU20A), PerCP-eFluor 710, eBioscience

Thermo Fisher Scientific BrdU Monoclonal Antibody (BU20A), PerCP-eFluor 710, eBioscience

Applications

• Flow Cytometry (Intracellular staining)

Tested Dilution: 5 µL (0.06 µg)/test
Publications: View 15 publications

Product Specifications

Additional Formats

• Unconjugated
• APC
• eFluor 450
• FITC
• PE
• Custom conjugation available

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 can identify cells that have undergone DNA synthesis during BrdU treatment.

• Recommended staining buffer sets:
  • BrdU Staining Buffer Set (Product # 00-5525)
  • Foxp3/Transcription Factor Buffer Set (Product # 00-5523)

Applications Reported: Intracellular staining followed by flow cytometric analysis.
Applications Tested: Pre-titrated and tested by flow cytometry on BrdU-pulsed mouse splenocytes.
Use at 5 µL (0.06 µg) per test (100 µL final volume, 10⁵–10⁸ cells/test).

Fluorochrome Information:
PerCP-eFluor® 710 can replace PE-Cy5, PE-Cy5.5, or PerCP-Cy5.5.

• Emission: 710 nm
• Excitation: 488 nm (Blue Laser)
• Recommended filter: 685 LP dichroic mirror and 710/40 band pass filter (695/40 acceptable)

Fixation Stability:
Stable with 2% formaldehyde overnight at 4°C. Evaluate for alternative fixation protocols.

BrdU Labeling and Staining Protocol (Summary)

1. Label dividing cells with 10 µM BrdU for 45 min at 37°C.
2. Wash once with 1X PBS.
3. Stain surface molecules per surface staining protocol.
4. Wash with cold Flow Cytometry Staining Buffer or PBS.
5. Resuspend and add 1 mL Foxp3 Fixation/Permeabilization Buffer (Product # 00-5521).
6. Incubate 30–60 min at 2–8°C in dark.
7. Wash and centrifuge; decant supernatant.
8. Resuspend in 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.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

Bromodeoxyuridine (BrdU) is a synthetic nucleoside analog of thymidine used to detect proliferating cells. Incorporated into newly synthesized DNA during the S phase, BrdU enables identification of actively replicating cells via anti-BrdU antibodies. Detection requires DNA denaturation (acid or heat treatment).

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

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