Bromodeoxyuridine Pulse Labelling Assay: A Technique to Measure Cell Proliferation
Published: April 30, 2023
Abstract
Source: Welschinger R. et al. Temporal Tracking of Cell Cycle Progression Using Flow Cytometry without the Need for Synchronization. J. Vis. Exp. (2015)
The video describes the stepwise pulse labeling assay of mammalian cells with bromodeoxyuridine (BrdU) for measuring cell proliferation. The BrdU uptake permits the temporal tracking of cells that were in the synthesis phase at a specific point in time, without requiring the cells to be synchronized.
Protocol
1. Solutions and Reagents Complete RPMI Add 56 ml fetal calf serum (FCS) and 5.5 ml of 200 mM L-glutamine to a 500 ml bottle of RPMI-1640 medium. BrdU Stock Solution Prepare 32.5 mM BrdU (10 mg/ml) in Dulbecco's Phosphate Buffered Saline (DPBS). BrdU Complete RPMI Add 6.2 µl of BrdU stock solution to 10 ml of Complete RPMI. DNase Solution Prepare 1 mg DNase/ml in DPBS. …
Açıklamalar
The authors have nothing to disclose.
Materials
| NALM6 | DSMZ | ACC-128 |
| 5-Bromo-2′-deoxyuridine | Sigma | B5002-1G |
| RPMI 1,640 w/o L-Gln 500 ml | Lonza | 12-167F |
| DPBS | Lonza | 17-512F |
| Fetal Bovine Serum | FisherBiotec | FBS-7100113 |
| CO2 incubator | Binder | C 150 |
| BD Falcon 12 x 75 mm FACS tubes | BD Biosciences | 352008 |
| BD LSR FORTESSA flow cytometer | BD Biosciences | FORTESSA |
| Centrifuge | Spintron | GT-175R |
Bu Makaleden Alıntı Yapın
Bromodeoxyuridine Pulse Labelling Assay: A Technique to Measure Cell Proliferation. J. Vis. Exp. (Pending Publication), e20258, doi: (2023).