Cheap and quick cell viability assay - (May/18/2006 )
QUOTE (haringsh @ Jun 30 2006, 01:20 PM)
The Promega kit that I described goes from cells to results in 10 minutes. The problem with Trypan Blue is that you have to count the cells within 5 minutes of adding the stain, otherwise you'll get false positives. While this isn't a problem if you're working with one or two cell lines, trying to do anything high-throughput can prove extremely tedious.
Before trying the Promega assay I did TB counts for 4 different mutants in triplicate, but I could only stain and count one mutant each time without passing that 5 minute mark (I also do my counts in triplicate).
TB is definitely cheaper and easier if you're not doing anything in high volume. I suggest reading this paper if you're looking at an alternative to TB or some commercial kit:
http://www.ncbi.nlm.nih.gov/entrez/utils/l...sue=6&spage=693
I haven't had the opportunity to try this out yet, but it looks promising.
Hank
Before trying the Promega assay I did TB counts for 4 different mutants in triplicate, but I could only stain and count one mutant each time without passing that 5 minute mark (I also do my counts in triplicate).
TB is definitely cheaper and easier if you're not doing anything in high volume. I suggest reading this paper if you're looking at an alternative to TB or some commercial kit:
http://www.ncbi.nlm.nih.gov/entrez/utils/l...sue=6&spage=693
I haven't had the opportunity to try this out yet, but it looks promising.
Hank
Thanks. I'll check this out.
Casey
-casandra-
You can use cristal violet (1 to 2 % in H2O) for ahderent cell in 96well plate. You remoove medium, add cristal violet solution to the cells for ca 20 min and wash intessive in a big volumen of H2O. Than add DMSO and read by 550nm. Cristal violet binds to proteins. and more proteins=more cells =higher OD. I have good expirence with crisal violet better than with MTT ( you do not nedd 4 h of incubation).
Hou use also cristal violet?
-LUK-