To study the proliferation of HSPCs, mice were injected with 10 mg/ml 5-brom-2′-deoxyuridin (BrdU, Sigma Aldrich, # B9285-250MG) as early as 1 hour before harvest. BM cells were obtained and prepared for flow cytometric analysis as described above. To analyze the proportion of BrdU in the cell populations, and therefore proliferating cells, further staining was performed after the primary antibody staining. Since BrdU is incorporated into DNA and is therefore an intracellular marker, cells were fixed and permeabilized using a Fix & PERM™ cell permeabilization kit (Invitrogen, #GAS004). The cells were then treated with deoxyribonuclease (DNase) to release the BrdU incorporated into the DNA. Cells were then stained with PE anti-BrdU antibody (Biolegend, San Diego, USA, #364116). Proliferating cells were identified as BrdU positive (BrdU+).
Pe anti brdu antibody
Manufactured by BioLegend
Sourced in United States
PE anti-BrdU antibody is a fluorescently labeled antibody that binds to bromodeoxyuridine (BrdU), a synthetic nucleoside that can be incorporated into newly synthesized DNA as a marker for cell proliferation. This antibody is designed for flow cytometry applications to detect and quantify BrdU incorporation in dividing cells.
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2 protocols using pe anti brdu antibody
1
Quantifying HSPC Proliferation with BrdU
2
Quantifying Cardiac Cell Proliferation
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To assess the proliferative activity of CD45negc-kitpos cardiac cells, before harvest in culture, BrdU (50 mg/kg) was intraperitoneally administered in vivo for 6 days in adult mice every 12 h, before mouse killing, and then it was administered in the enzymatic solution (10 μM) while digesting the hearts. After harvest in culture BrdU 10 μM was administered in vitro every 12 h for 7 days on freshly isolated CD45negc-kitpos cardiac cells and CD45negc-kitpos cardiac cells at P1. Then cells were analysed by flow cytometry (PE Anti-BrdU Antibody, Biolegend, San Diego, CA, USA).
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