Thank You for sharing this protocol : I have 4 questions
1- How could I determine the PH in my reaction? I do the same volumes mentioned in the kit which differ according to the plate size (usually do 10 cm petri dish , without assessing the PH) I didn`t get proper greenish bluish staining , usually more brownish , I incubate them for 24 hours in dry 37 degree incubator without co2 as recommended.
2- My treatment is UV irradiation , do you have any idea how long should I keep cells after UV ttt to be able to attain senescence? in some paper they left them up to 12 days which is hard for my cells to be left all this time without need to be passaged?
3- What is the recommended initial number or confluency of the flask prior to ttt if you intend to keep cells 5 days for ex. before fixation and staining?
4- I use the fixative solution that come with the kit and it is mentioned to be kept for 12-15 min. before washing, Is this different than paraformaldehyde 4% you are using?
1/7/2020 6:05:08 AM Reply