Help-apoptosis of adhesion cell
Posted 27 March 2005 - 01:26 AM
Thanks a lot!
Posted 29 March 2005 - 10:59 AM
Posted 31 March 2005 - 01:44 AM
I used to do Annexin V staining for FACS on adherent cell lines. I found that my trypsinisation using commercial mix of trypsin/EDTA only resulted in a very small percentage of damaged cells <1%. I guess that it is vastly dependant on the cell type.
Have you considered staining the cells on cover slips and examining by IF and just doing cell counts?? Or fixing cells prior to harvesting, not sure if either would work but could try. The other possiblity is that there is any number of commercial kits these days for assaying apoptosis you could check these out.
Sorry I haven't been much help.
Posted 31 March 2005 - 04:32 PM
If you are getting high background you may need to add a blocking agent such as 1% BSA or IgG to your buffer. Also, using propidium iodide you can tell if your cells are being damaged from your treatment (removal from the flask).