Hi Preeti,
Thank you for reading our protocol. These SNs can be used in all of the applications you mentioned; however, if you store them at -80, you will compromise the SNs integrity. If you want to perform functional and structural studies, you should use fresh SNs. Additionally, the EGTA in the buffers can be decreased or removed to reduce the interference with calcium signaling (the protease activity will increase too).
Best,
Ariel
2021-02-19 06:16:35