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

Play Video

Bu Makaleden Alıntı Yapın
Bromodeoxyuridine Pulse Labelling Assay: A Technique to Measure Cell Proliferation. J. Vis. Exp. (Pending Publication), e20258, doi: (2023).

View Video