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[miss]

I'am doing a time course for apoptosis......with my drug I can see caspase, parp cleavage and modulation of other apoptotic markers in as early as 6hrs but when I do apoptotic staining with AnnV/PI stain I can see no staining in 6,12hrs very little AnnV /PI staining (5%) in 24hrs and around 20 % in 36hrs.............why is it so since I see the proteins modulated in as early as 6hrs.............Also can some one let me know what is the sequence of apoptotic events specially at what point PS exposure occurs

[Stephan]

As far as I remember the major events are (in order)

• PS exposure
  
• Caspases
  
• Cytochrome C
  
• DNA cleavage