Luciferase: too much pbs - (Aug/08/2012 )
If I double the amount of PBS per well of cells that a luciferase protocol calls for, should I double the reporter lysis buffer as well? Do I then add twice as many cells to my plate when I set it up for reading on the luminometer?
It depends on what the roe of the PBS is? - do you remove it before adding the lysis buffer? if so, it is a wash step and you can proceed without doubling.
No, it's not removed. Lysis buffer is added directly.
If I double the amount of lysis buffer, do I also double the amount of assay and substrate I use (200ul instead of 100ul)?
Probably, as you will need to maintain a minimum concentration for those reagents.