BrdU/GFP/PI labeling
ZBIGNIEW DARZYNKIEWICZ (darzynk@nymc.edu)
Thu, 04 Jul 1996 09:45:53 -0500
Dr. Wentraub inquired about a method which may allow to
simultaneously detect the incorporated BrdUrd and be gentle enough
not to destroy GFP. The SBIP (DNA Strand Breaks Induction by
Photolysis) approach may do the trick. In this method, after BrdUrd
incorporation, the cells can be fixed or permeabilized in any desired
way, then subjected to UV illumination, and DNA strand breaks induced
by photolysis (at the sites of BrdUrd incorporation) labeled with
BrdUTP, which in turn is detected by FITC-anti BrdUrd-MoAb (Cell
Proliferation, 28: 571-579, 1995). There is no DNA denaturation step,
which is harsh and generally impairs the immunocytochemical detection
of other cell constituents immunocytochemically.
Zbigniew Darzynkiewicz