lithium chloride - wnt/bet catenin pathway (Apr/14/2011 )
Hi,
Does anyone have experience in tearting cells with LiCl to study the wnt/beta catenin pathway. I need to treat my cell lines with various concentration of LiCl. I have been to incubate teh cells with 20mM LiCl prepared in water. My questions are:
1) If I prepare a stock of 0.5M LiCl in water and dilute it further with water to give a concentration of 20mM, is it on to incubate cells with LiCl dissolved in water?
2) Do I have to serum starve the cells prior to incubating them with LiCl.
Thank you!!
I have treated cells with LiCl to look at reduction in GSK3 activity and what i do is to make a 5M stock of LiCl in water (filter sterilize). Then I dissolve that stock solution into media. I have never had to serum starve cells first.
kfunk106 on Thu Apr 14 17:39:53 2011 said:
I have treated cells with LiCl to look at reduction in GSK3 activity and what i do is to make a 5M stock of LiCl in water (filter sterilize). Then I dissolve that stock solution into media. I have never had to serum starve cells first.
Hi, thank you for your helpful response. I had prepared 0.5M LiCl in water, filter sterilised and then diluted that with cell culture medium. I'm curenty testing 20mM and 40mM concentrations for 12hr, 24hr and 48hr time periods. Can I ask at what confluency did you treat the cells with LiCl. My cells were 80-90% confluent at the time of treatment. I've got very limited time to finish this experiment and all advice will be appreciated.