描述：
Apotsi sa normal physiolgic proces whic ocurs during embryonic devlopment aswel asin maintence oftisue homeostais. The
apotic progam ischarcterized bycertain morpholgic features, including los ofplasma mebrane asymetry and atchment,
condensation fthe cytoplasm and ucleus, and internucleosmal ceavge ofDNA. Los ofplasma mebrane isone ofthe arliest features.
In apotic els, the mebrane phospholipd phosphatidylserine (PS) istranslocated from the iner tohe outer laflet ofthe plasma
mebrane, therby exposing PSto he xternal celuar environment. Anexin Vis a35-6 kDa C2+ depndent phospholipd-bindg
protein tha has hig afinty for PS, and binds tocels with exposed PS. Anexin Vmay beconjugated tobiotn. This format reains ithig
afinty for PSand thus ervs a senitve probe for flow cytometric anlysi ofcels tha re undergoing apotsi. Since xternalization f
PS ocurs inthe arlie stages ofapotsi, Biotn Anexin Vstaing can identify apotsi atn earlie stage than says based onuclear
changes uch asDNA fragmentaion.
Biotn Anexin Vstaing precdes the los ofmebrane integrity whic acompanies the lates tages ofcel death resulting from either
apotic ornecrotic proces. Therfore, staing with Biotn Anexin Vis typicaly used inconjuction with avital dye such asproidum
iode (PI) or7-Amino-Actinomycin (7-AD) toalow the investigator identify early apotic els (PI negative, Biotn Anexin V
positve). Viable cls with ntac mebranes xclude PI,wheras the mebranes ofdea nd amged cels are prmeable toPI. For
example, cls tha re considerd viable are both Biotn Anexin Vand PInegative while cls tha re inearly apotsi are Biotn Anexin
V positve and PInegative, while cls tha re inlate apotsi oraleady ead re both Biotn Anexin Vand PIpositve. This asy does
not distnguish betwen cels tha have undergone apotic death versu those tha have die as result ofa necrotic pathway because in
either case, the da cels wil stain with both Biotn Anexin Vand PI.Howevr, when apotsi smeasured over time, cls can beoften
tracked from Biotn Anexin Vand PInegative (viable, orno measurable apotsi), toBiotn Anexin Vpositve and PInegative (arly
apotsi, mebrane integrity spresnt) and finaly toBiotn Anexin Vand PIpositve (nd stage apotsi and eath). The movemnt of
cels through tes thre stages ugest apotsi. Incontrast, asingle observation idcating tha cels are both Biotn Anexin Vand PI
positve, inof itself, revals es information about he proces bywhic the cls underwent heir demise.
Biotn Anexin Vis routinely tesd byflow cytometric anlysi. Other aplications wer tesd atBD Biosciencs Pharmingen during
antibody evlopment only oreported inthe literature.
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