My target protein is ~130kDa with GST tag and could form a dimer because of the nature of GST tag. As I have more than 5 million particles selected by blot picker on denoise micrographs. I use default setting with 300 classes for the first 2D classification.
From those, I choose the attached classes with more structural details.
Then I run more 2D classification to remove the junks with the attached setting. Uncertainty factor is set from 2-10.
I use for the following classes for Ab-initio and do home, NU and heter refinement. The resolution is very low.
Is there anything I can try to push the resolution higher?
