Re: Apoptosis light scatter detection

From: Karim Vermaelen (Karim.Vermaelen@rug.ac.be)
Date: Tue Jun 18 2002 - 05:34:08 EST


Hi William,
I was particularly intrigued by the multiple simultaneous labeling of
caspases you mentioned in your last message... Do you have any reference or
protocol description? Which manufacturer makes the best tools?
Thanks!
Karim

On 14/6/2002 6:31, "William Telford" <TelfordW@mail.nih.gov> wrote:


>
> (1)  If you use fluorogenic caspase substrates to label multiple caspases
> simultaneously (such as 8 and 3), the negative, single-positive and
> multiple-positive apoptotic subpopulations all have slightly differing
> scatter properties.  If looked at sequentially (like 8-3-, 8+3- and 8+3+,
> for example), the cell size appears to increase slightly during proximal
> caspase activation and return to "normal" after distal caspase
> activation.  This is of course all much later than the phase of cell death
> associated with mitochondrial membrane potential flux, when the cell volume
> changes described by you are timed to occur.  However, they are still much
> earlier than DNA dye permeability and annexin V binding.  And this is MUCH
> earlier than the gross changes in cell size associated with late cell death.
>



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