Detection of Bromodeoxyuridine/BrdU in Human PBMCs by Flow Cytometry.
Human peripheral blood mononuclear cells (PBMCs) were treated overnight with 50 ng/mL PMA, 500 ng/mL Ionomycin, and 30 µg/mL BrdU, then stained with Mouse Anti-Bromodeoxyuridine/BrdU APC‑conjugated Monoclonal Antibody (Catalog # IC7225A, filled histogram) or isotype control antibody (Catalog # IC003A, open histogram). To facilitate intracellular staining, cells were fixed with cold, 70% ethanol for 5 minutes, DNA was denatured with 1.5M HCl for 30 minutes, and then cells were permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005).View our protocol for Staining Intracellular Molecules.
Preparation and Storage
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Protect from light. Do not freeze.
12 months from date of receipt, 2 to 8 °C as supplied.
Bromodeoxyuridine (BrdU) is a nucleoside analog that is incorporated into DNA in place of thymidine. BrdU differs from thymidine by virtue of the fact that BrdU shows a bromine atom substitution for a methyl group in thymidine. The detection of newly synthesized DNA containing BrdU is a commonly used measure of cell proliferation and progression through S phase of the cell cycle.
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