Hi

Could you clarify the question, what is the luciferase measuring -is it extracellular ATP or intracellular? If the assay is for ATP release, won't the extracellular BzATP be constant at (say) 0.1. mM once you apply it so any extra ATP release will be seen as a second slower increase? What is the concentration of agonist (BzATP) compared to what the luciferase is expected to report?

HTH M

Good question Jordan. I have never thought of it before. I will ask other researchers that work with BzATP.

Thank you for your responses, Mark and Ana!

I would be using BzATP at around a in the uM range and measuring intracellular ATP at an expected mM range. The assay will be to measure intracellular ATP using not luciferase, but another biological ATP-binding ligand, with BzATP being applied extracellularly. Since I know some biological ligands (e.g., P2X7) bind both ATP and BzATP, I am interested if others do as well (e.g. luciferase).

I may just have to do a luciferase assay and see empirically what's going on. If I find out before someone can answer, I'll report back.

It is an interesting problem since ATP is tested frequently in the present of BzATP.

Here is a reference that would validate it: Mechanisms of constitutive and ATP-evoked ATP release in neonatal mouse olfactory epithelium Sébastien Hayoz, Cuihong Jia, CC Hegg BMC Neuroscience 2012, 13:53 (28 May 2012)

However I heared that luciferase assay can measure bzatp. So actually an experimental affirmation of it would be very helpfull

What was the results of your tests?