PBMCs were irradiated with 60 Gy and cultivated at a concentration of 25 × 106 cells for 24 h. Cell cycle analysis was performed using the BrDU cell-cycle kit (BD Biosciences, Franklin Lakes, NJ, USA) according to the manufacturer's instructions. Briefly, 20 h after irradiation, cells were incubated with BrdU (10 µM, kit component) for 4 h. After fixation (BD Cytofix/Cytoperm™, kit component) cells were stained with a fluorescein isothiocyanate (FITC) conjugated anti-BrdU antibody (BD Biosciences, kit component) for 30 min. After washing, cells were stained with 7-AAD (BD Biosciences, kit component) and immediately analyzed on a FACS-Calibur (BD Biosciences). Gates were set according to the manufacturer's instructions, and data were evaluated using the FlowJo software (Tree Star, Ashland, OR, USA).
Brdu cell cycle kit
Manufactured by BD
Sourced in United States
The BrDU cell-cycle kit is a laboratory reagent used to analyze the cell cycle. It detects 5-Bromo-2'-deoxyuridine (BrDU) incorporation, which is a measure of DNA synthesis and cell proliferation.
Automatically generated - may contain errors
2 protocols using brdu cell cycle kit
1
Cell Cycle Analysis of Irradiated PBMCs
2
Cell Cycle Analysis via BrdU Staining
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Cell cycle analysis was performed using the BrdU cell-cycle kit (containing the reagents below; BD Biosciences, Franklin Lakes, NJ) according to the manufacturer's instructions. 48 h post transfection, cells were incubated with BrdU (10 µM) for 4 h. After fixation with 100 μL of BD Cytofix Cytoperm Buffer for 15 min at 4 °C cells were stained with a fluorescein isothiocyanate (FITC) conjugated anti-BrdU antibody for 30 min and stained with 7-AAD and immediately analyzed on a FACS-Calibur (BD Biosciences). Gates were set according to the manufacturer's instructions and data were evaluated using FlowJo software (Tree Star, Ashland, OR).
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