I started to use RossetteSep for my NK cell isolation with slight modification of manufacturer's instruction as follows:-

Manufacturer's instruction: RossetteSep is added into the whole blood directly (50ul/ml blood)

My modification:

Whole blood --> plasma extracted --> RBC pallet (after resuspend with pipette) was mixed with RossetteSep cocktail --> manufacturer's guidelines followed.

Suggested modification by superior:

Whole blood --> plasma extracted --> RBC pallet mixed with PBS --> RossetteSep cocktail added--> manufacturer's guidelines followed.

Any suggestion?