Dynamics of membrane translocation of phosphatidylserine during apoptosis detected by a monoclonal antibody.
Translocation of phosphatidylserine (PS) to the outer leaflet of the cellular membrane seems to be a key step in apoptosis and cell activation. In this paper, the production and characterization of a monoclonal antibody designated as Mab 1H6 is described which does not show cross reactivity with others anionic phospholipids. It is demonstrated that Mab1H6 can recognize externalized PS at early stages after the induction of apoptosis shown by both flow cytometry and immunofluorescence. Our results show that translocation of PS can be detected as early as 5 min by immunofluorescence and 10 min by flow cytometry after the treatment of cells and a specific dynamics is observed concerning the location and distribution of the staining. These data prove that antibody Mab 1H6 can be used as a specific probe for detection of PS translocation.
Mourdjeva
M
M
Institute of Biology and Immunology of Reproduction, Bulgarian Academy of Sciences, Sofia, Bulgaria.
Kyurkchiev
D
D
Mandinova
A
A
Altankova
I
I
Kehayov
I
I
Kyurkchiev
S
S
D000911:Antibodies, Monoclonal / Q000378:metabolism*; D017209:Apoptosis / Q000187:drug effects*; D002462:Cell Membrane / Q000276:immunology / Q000378:metabolism*; D053938:DNA Fragmentation; D005434:Flow Cytometry; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D019169:Jurkat Cells; D018613:Microscopy, Confocal; D016877:Oxidants / Q000494:pharmacology*; D010718:Phosphatidylserines / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D016877:Oxidants; D010718:Phosphatidylserines; D006861:Hydrogen Peroxide