Re: annexin V/apoptosis

From: Eric Miller (millere@icrf.icnet.uk)
Date: Mon May 08 2000 - 03:51:34 EST


Hi Rena
	We routinely use FL1-H versus FL2-H, both log. The only advantage
I can see in using FL3 is that because of the wider separation of bandpass
filters compensating out crossover would be less of a problem
	To your other query, we routinely use adherent cells in our
annexin work. We find no problem in using trypsin/EDTA to detach cells,
providing you return your cells to the incubator suspended in complete
medium to allow the cells to recover from the detaching. It is important
to handle the cells very gently: the assay depends absolutely on intact
cell membranes. One point we have recently become aware of: don't vortex
your cells too violently before running samples. We have seen some very
odd results due to this!

	Eric P Miller
	Edinburgh Medical Oncology Unit

"Everyone can be correct, all at the same time. That's the
 thing about quantum."
	Terry Pratchett, Lords and ladies

On Wed, 3 May 2000, feinmarr wrote:

>
> Hi:
> I need your input.
>
> What are the best parameters to use when acquiring samples for
> FITC-annexinV and PI?  In the past, our parameters were FL2-H vs FL1-H.
> My colleague prefers using FL3-H vs FL1-H. What are the pros and cons
> for both ways? We are using myeloma cell lines.
>
> My second question: can you perform annexin V binding on adherent cell
> lines?
> Thanks in advance, Rena
>
>
>
>
>
>



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