Here are some suggestions for dissolving lyophilized actin powder in insoluble droplets.

Use an incremental buffer.

Lyophilized actin powder may not dissolve completely in the same amount of buffer originally used for reconstitution.

Try using a larger buffer like 2x or 3x, and see if this helps.

Use a different buffer.

Lyophilized actin powders may be more miscible in a different buffer than those originally used for reconstitution.

Try using a buffer with a different pH or ionic strength.

Heat the protein solution.

Heating a protein solution can help degrade the protein and make it more soluble.

Try to heat the protein solution to 37°C or 42°C for 10-15 minutes.

Use mild detergent.

Mild detergents such as Triton X-100 or Tween 20 can help dissolve proteins by disrupting protein-protein interactions.

Try adding a small amount of detergent (0.1-0.5%) to the protein solution.

Use a sonicator.

A sonicator can be used to disrupt protein aggregates and make proteins more soluble.

Try to sonicate the protein solution for 1-2 minutes.

If none of these suggestions work, you can try a stronger approach, such as using a laxative such as urea or guanidine hydrochloride.

However, this can damage the protein and make it inactive.

Here is a specific protocol you can try:

Dissolve the lyophilized actin powder in a small amount of buffer (e.g., 100 µL).

Add a small amount of detergent (e.g., 0.1% Triton X-100).

Soak the protein solution in water for 1-2 minutes.

Heat the protein solution to 37°C or 42°C for 10-15 minutes.

Vigorously vortex the protein solution.