Thank you in advance.
I need to make my internal that I'm going to use but I need some guidance. I will be making my internal in two parts. The first part will consist of the buffer & salts, while the second part will consist ATP & GTP. My plan is to mix 900uL of the first part and 100uL of the second part the day of recording. The pH that has been traditionally used in the lab is 7.3.
I calculated what the concentration needs to be for each of the internal components so that when combined will yield the desired 'working' concentration.
However, my questions are the following: How am I supposed to pH two part internal recipes? Do I only pH the major constituent (buffer and salts) and not worry about the effect of the ATP/GTP on the pH)? What about osmolarity? Do the same?
I am the only one in the lab doing e-phys and now its time for me to start doing whole cell work.
Once again thank you in advance,