Tuesday, August 21, 2007
Flow cytometry_GFP and PI or DNA simotaneously
A good method to fix cell to detect GFP signal:
Fixation reagent:
2% Paraformaldehyde, 100 mM NaCl, 300 mM Sucrose, 3 mM MgCl2, 1mM EDTA, 10 mM PIPES, pH 6.8
Dyhydration : 70 % ice-cold ethanol
Procedure:
wash cells with PBS, then Fixation reagent for 30 min, room temperature, PBS wash 4X, then 70% ethanol for 10-12 hours. PBS wash, RNaseA (50ug/ml) for 30 min, PBS wash, PI (50ug/ml) 30 min before FACS analysis.
Reference:
Nucleic Acids Research, V. 25, p 4855-4857.
A rapid, quantitative and inexpensive method for detecting apoptosis by flow cytometry in transiently transfected cells.
Lamm GM, Christofori G.
Fixation reagent:
2% Paraformaldehyde, 100 mM NaCl, 300 mM Sucrose, 3 mM MgCl2, 1mM EDTA, 10 mM PIPES, pH 6.8
Dyhydration : 70 % ice-cold ethanol
Procedure:
wash cells with PBS, then Fixation reagent for 30 min, room temperature, PBS wash 4X, then 70% ethanol for 10-12 hours. PBS wash, RNaseA (50ug/ml) for 30 min, PBS wash, PI (50ug/ml) 30 min before FACS analysis.
Reference:Nucleic Acids Research, V. 25, p 4855-4857.
A rapid, quantitative and inexpensive method for detecting apoptosis by flow cytometry in transiently transfected cells.
Lamm GM, Christofori G.
# posted by yuanchin : 10:31 AM
