Ammonium sulfate precipitation of proteins protocol

The following protocol is for secreted proteins. Modify according to other soluble proteins

  1. The spin down cells
  2. Pour the supernatant into a flask or a glass
  3. Remove the small part to analyze the gel
  4. Add the desired amount of ammonium sulphate very slowly (in the form of a saturated liquid, or in powder form)
  5. Stir for 1 hour
  6. Spin down protein deposited in 10,000 x g., 30 min
  7. Resupend pellets in PBS for dialysis (concentrates approximately 1 / 6x to 1 / 10x based on the amount of the original media)
  8. Take some supernatant from spinning down proteins for dialysis

desirable

Ammonium sulfate precipitation of proteins protocol
% Saturation The amount of saturated ammonium sulfate to add (to the media 10 ml)
10 1.11ml
20 2.5ml
30 4.29ml
40 6.67ml
50 10ml
55 12.22ml
60 15ml
65 18.57ml
70 23.33ml
75 30ml
80 40ml
85 56.66ml
90 90ml
95 190ml