I am working with GZMA/Granzyme A recombinant protein (HY-P76377 from MCE) in lyophilized powder form. After reconstituting it with 100 µL of ddH₂O and subsequently added 400µL of buffer containing carrier protein to obtain a final concentration of 100 µg/mL, I aliquoted 25 µL into tubes and stored them at -80°C.

I would like to know if in vitro activation is necessary for this protein prior to use. If so, could anyone provide an activation protocol based on my reconstitution process? Specifically, I am looking for details on the following:

Buffer requirements for activation: What specific buffer and formula should be used?

Activation enzyme information: Which enzymes are recommended for activation, what concentrations should be used, and what is the appropriate order for adding samples?

Activation conditions: What temperature and duration should be employed, and what specific steps should I follow?

Post-activation considerations: Is it necessary to remove the activation enzyme after the process?

Storage and stability: What are the best practices for storing activated proteins to maintain their activity?

Your insights and recommendations would be extremely valuable. Thanks in advance!