Detection of Apoptotic cells via FACS

5 posts / 0 new
Last post
Guy Sovak
Guy Sovak's picture
Detection of Apoptotic cells via FACS

Hi All,
Here is a link to detect Apoptotic cells using FACS method and three different Ab's: Hoechst, TUNEL and combination of Annexin V and 7-AAD.
http://biochem.dental.upenn.edu/~sdmfacs/apopproc.htm
This link comes from the:University of Pennsylvania
School of Dental Medicine

Links:
Here are some more links for those who dose not know about:
Hoechst: http://en.wikipedia.org/wiki/Hoechst_stain
7-AAD: http://www.ebioscience.com/ebioscience/specs/antibody_00/00-6993.htm
Annexin: http://www.bdbiosciences.com/pharmingen/protocols/Annexin_V_Staining.shtml (this is also a protocol for the staining of Annexin V.

samm
samm's picture
Sadly the Hoechst dyes do not

Sadly the Hoechst dyes do not work in most flow cytometers - unless the always "optional eq" upgrade to the very expensive ~300 nm uv laser is done. Annexin-FITC with PI or Annexin-PE with 7-AAD does work like a charm though (as long as you are careful with the compensation!).

marcus muench
marcus muench's picture
The recent marriage of

The recent marriage of molecular probes with Caltag and Invitrogen has resulted in a number of new options for live/dead cell and apoptosis staining. (Maybe not new, but at least yanked from the obscurity that was the molecular probes catalog and marketed better.)

Looky here for anyone that may be interested:
http://probes.invitrogen.com/servlets/publications?id=608&mode=list

Note, the SYTOX Red dead cell stain is suggested as a good combination for Annexin V-PE. I haven't tried it myself, but it is on order. I'm alsoo looking at the violet laser options. It would be nice to free up the "FL-3" channel by replacing PI.

The only problem with most of the newer offerings is that they are very expensive compared to the relatively cheap PI and 7-AAD.

I heard a rumor that DAPI works with violet lasers. Has anyone tried using this dye on one of the newer flow cytometers?

wyzandrea
wyzandrea's picture
Hi,
susanwn
susanwn's picture
 as for the apoptosis, I also

 as for the apoptosis, I also learn something about it. Once I have used the FACS method to detect apoptotic cells. it is really useful. thanks for sharing so many methods here.