This video describes the protocol for BrdU immunofluorescence staining. The technique allows the identification of cells in different phases of cell cycle. Addition of DNA dyes and antibody labeling allows the detailed analysis of the fate of the S phase cells at later times.
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. DNase Solution Prepare 1 mg DNase/ml in DPBS. Staining Buffer Prepare 3% heat-inactivated FCS and 0.09% sodium azide in DPBS. Refer to Materials List for definitions of Fixation Buffer, Permeabilization Buffer and Wash Buffer. …
Açıklamalar
The authors have nothing to disclose.
Materials
APC BrdU Flow Kit
BD Biosciences
552598
Contains BrdU antibody, 7-AAD and BD Cytofix/Cytoperm Buffer (referred to as Fixation Buffer)
BrdU Immunofluorescence Staining: A Technique to Identify Cells in Different Phases of Cell Cycle. J. Vis. Exp. (Pending Publication), e20259, doi: (2023).