Sigma Aldrich, USA
PKH67 Green Fluorescent Cell Linker Mini Kit for General Cell Membrane Labeling
MINI67-1KT
Aim of the present study was to fluorescent labeling of pancreatic beta cells plasma membrane of cells treated with or without glucose in presence or absence of insulin
Membrane labeling of rat pancereatic beta cells in vitro
Rat pancreatic beta cell culture, Trypsin EDTA, Trypsin neutralizer, DMEM with serum, DMEM without serum, PKH67 Green Fluorescent Cell Linker Mini Kit, DPBS or HBSS, cell culture incubator, centrifuge, etc.
Rat pancreatic beta cell suspension culture maintained at suitable environment was set to a cell concentration of 107/ ml. Cells were washed two times with serum free medium and supernatant were discarded remaining cell suspension volume up to 25 micL. Cells were then re-suspended up to 1 ml in diluents C (supplied with kit) to make 2x cell suspension. Further, above cell suspension was slowly added to 1 ml of 2x dye solution (4 micro molar) which make 2 micro molar PKH67 green fluorescent cell linker suspension. Cells were incubated for 3 minutes with periodic mixing and linking reaction was stopped by adding equal volume of serum containing medium. At end cell suspension was centrifuged at 400 xg for 5 min and seeded into suitable culture dishes.
None
There were labeling of plasma membrane of pancreatic beta cells as detected under fluorescent microscope. Labeling steps were so easy and labeling persists up to 24 hrs.
N/A
Easy method of plasma membrane labeling
Kit is costly
Good kit for plasma membrane labeling