Cell cycle analysis was performed using the APC‐BrdU Flow Kit (BD, Catalog No.: 552598). Coculture cells or cord blood total nucleated cells were treated with BrdU for 6 h before analysis. All cells were first stained for surface Ags, and then processed using an APC‐BrdU Flow Kit (BD) according to the manufacturer's instructions. All cells were detected using a FACSCanto II system (BD Biosciences).
Apc brdu flow kit
Manufactured by BD
Sourced in United States, France
The APC BrdU Flow Kit is a laboratory equipment product that enables the detection and measurement of bromodeoxyuridine (BrdU) incorporation into cellular DNA during DNA synthesis. It provides the necessary reagents and protocols to perform this analysis using flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Bromodeoxyuridine (brdu), by Merck Group (13 mentions)
Bromodeoxyuridine (brdu), by BD (11 mentions)
Facscanto 2, by BD (11 mentions)
Facscalibur, by BD (10 mentions)
Facscanto, by BD (7 mentions)
Facscanto 2 flow cytometer, by BD (6 mentions)
Cytofix cytoperm buffer, by BD (6 mentions)
Facscalibur flow cytometer, by BD (6 mentions)
Attune flow cytometer, by Thermo Fisher Scientific (5 mentions)
Pe annexin 5 apoptosis detection kit, by BD (5 mentions)
Lsrfortessa, by BD (5 mentions)
Lsrii flow cytometer, by BD (5 mentions)
Dapi staining, by Merck Group (5 mentions)
Moflow, by Beckman Coulter (4 mentions)
Lsr 2 flow cytometer, by BD (4 mentions)
Fortessa flow cytometer, by BD (4 mentions)
Facsdiva software, by BD (4 mentions)
Annexin 5 apc, by Thermo Fisher Scientific (3 mentions)
Celltrace violet, by Thermo Fisher Scientific (3 mentions)
Facsaria, by BD (3 mentions)
275 protocols using apc brdu flow kit
1
Cell Cycle Analysis Using APC-BrdU Flow Kit
2
Cell Cycle Analysis of HeLa Cells
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HeLa cells were grown in 6-well plates (2 × 105 cells/well) for 12 h and then straved without serum for 16 h, treated with PBS or rLm-PHB2 at 0.625 μM, 1.25 μM and 2.5 μM for 24 h. BD Biosciences APC BrdU Flow kit (cat # KGA319-03) was used for analysis. BrdU (10 uM) was added to culture medium and cells were incubated for an additional 4 hours, then harvested with 0.25 % trypsin and 0.1 % EDTA in HeLa cells and fixed and stained according to the protocol provided by the BD Biosciences APC BrdU Flow kit and analysized on flow cytometer. Analysis was performed using Flow jo 7.6.1 software.
HeLa cells were grown in 6-well plates (2 × 105 cells/well) for 12 h, and then transfected with pEGFP-N1 or pEGFP-N1-Lm-PHB2 plasmid for 24 h. After removing the medium, the adherent cells were digested with ethylene diaminetetraacetic acid (EDTA) free trypsin (HyClone, USA) and harvested by centrifugation. The cells were then washed twice with PBS and fixed in 500 μL ice-cold 70 % ethanol at 4 °C for 2 h. The fixed cells were then centrifuged at 2000 × g for 2 min and the pellet was washed with PBS. After that, the cells were resuspended in 100 μL RNase I and incubated at 37 °C for 30 min followed by the addition of 400 μL propidium iodide and further incubation at 4 °C in the dark. The sample was finally analyzed by flow cytometry.
HeLa cells were grown in 6-well plates (2 × 105 cells/well) for 12 h, and then transfected with pEGFP-N1 or pEGFP-N1-Lm-PHB2 plasmid for 24 h. After removing the medium, the adherent cells were digested with ethylene diaminetetraacetic acid (EDTA) free trypsin (HyClone, USA) and harvested by centrifugation. The cells were then washed twice with PBS and fixed in 500 μL ice-cold 70 % ethanol at 4 °C for 2 h. The fixed cells were then centrifuged at 2000 × g for 2 min and the pellet was washed with PBS. After that, the cells were resuspended in 100 μL RNase I and incubated at 37 °C for 30 min followed by the addition of 400 μL propidium iodide and further incubation at 4 °C in the dark. The sample was finally analyzed by flow cytometry.
3
BrdU Incorporation Assay for HSCs and MPPs
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Mice were injected with a single dose of 5-bromo-2′-deoxyuridine (BrdU; 1mg/10g body mass) and maintained on 1 mg BrdU/ml drinking water for three days. Then HSCs or MPPs were isolated as described above, fixed, and stained using the BrdU APC Flow Kit (BD Biosciences), and analysed for BrdU incorporation. For the other cell populations analysed, which are more abundant than HSCs and MPPs, cells were stained with cell surface antibodies, then fixed and stained using the BrdU APC Flow Kit (BD Biosciences) and analysed for BrdU incorporation.
4
BrdU Incorporation Assay for HSCs and MPPs
5
Cell Growth and Cycle Analysis of PRDX6 Knockout
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SNU475PRDX6+/+ and SNU475PRDX6−/− cells (3.5 × 104) were seeded in 24-well plates and allowed to grow to generate growth curves of counted cells at 24, 48, and 72 h. For cell cycle analysis, 5 × 105 of these cells were seeded in a 60 cm2 plate and incubated for 48 h. After that, a 3 h pulse with BrdU (10 mg/mL) was carried out for BrdU incorporation, as determined using the APC BrdU Flow Kit (Becton Dickinson Biosciences, Franklin Lakes, NJ, USA). The proportions of cell cycle phases were determined using flow cytometric analyses of 7-AAD-stained SNU475 cells. The cytometer used was the BD AccuriTM C6 Plus (BD Biosciences). The data were processed with BD Accuri™ C6 Plus 1.0.34 software (BD Biosciences).
6
Cell Cycle Analysis by BrdU Flow Cytometry
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The APC BrdU Flow Kit (Becton Dickinson, Franklin Lakes, NJ, USA) was used for flow cytometric analysis of the cell cycle according to the manufacturer’s protocol. After the respective treatment (either cytokines, drugs, or controls), BrdU solution was added for another 3 h into the medium of the cells (10 µL of 1 mM BrdU per mL culture medium). Then, both the adherent as well as the detached cells in the supernatant were collected and counted. A total of 5 × 105 cells/well were placed into a 96-deep-well plate, fixed, permeablized, and stained according to the manufacturer’s protocol. Flow cytometric measurements were performed on an LSRII cytometer in combination with the FACSDiva software V. 9.0, and the derived data were analyzed using the FlowJo software V. 10.8.0 (all from Becton Dickinson).
7
Cell Proliferation Analysis by BrdU Incorporation
8
Cell Cycle Analysis by Flow Cytometry
9
Quantifying Cell Proliferation Using BrdU
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To identify proliferating cells, NPPC were expanded for 7 days in proliferation medium (alpha minimum essential medium (α-MEM; Gibco, Life Technologies) containing 10 % fetal bovine serum (FBS; Sigma-Aldrich) and penicillin/streptomycin (P/S, 100 units/ml and 100 μg/ml, respectively; Merck, Darmstadt, Germany)), whereby 10 μM bromodeoxyuridine (BrdU) was added at the beginning of the experiment with one medium change. The incorporated BrdU was detected by flow cytometry according to manufacturer’s instructions (APC BrdU Flow Kit; Becton Dickinson).
10
Assessing Cell Cycle Dynamics
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