As long as standards are assigned first, controls and unknowns will automatically be quantified from the standard curve. However, MSD Discovery Workbench software will report unknown samples as N/A or NaN if the unknowns did not have standards assigned to them in the plate layout. In this case you will need to:
1. Return to the plate layout
2. Highlight the unknowns and select the box 'U' at the top. The 'Assign Unknowns to wells' window will open.
3. Make sure that the unknowns are back-fitted to the corresponding standard.
This can be applied to controls as well.