Question

Exercise 5

CHM 222 Exploratory Assignment Name: Score: Exercise 5: After the protein is purified in a column, the pro dialysis buffer is prepared as followed: 50 mM Potassium Buffer Protein is purified in a column, the protein has to be dialyzed in an appropriate buffer. The prepared as followed: 50 mM Potassium Buffer (KH,PO4 +K HPO.) stored as a lM solution, 0.5 mM EDTA stored has a 1 M solution, and 150 mM Na stored has a 1 M solution, and 150 mM NaCl stored as a IM solution. The total volume needed for dialysis is 2L culate the volume in mL of Potassium Buffer, EDTA, and NaCl needed to be added to the solution b) Describe how the solution would be prepared in 2-3 steps.

Answer 1

(a)

After mixing these solutions , total volume becomes 2L i.e. 2000ml

We can apply M1V1 = M2V2

1M× V_buffer = 50×10^-3M × 2000ml

V_buffer = 100ml

1M×V_NaCl = 150×10^-3M×2000ml

V_NaCl = 300ml

1M×V_EDTA = 0.5×10^-3M× 2000ml

V_EDTA = 1ml

Volumes of potassium buffer , EDTA and NaCl needed to form the solution = 100ml, 1ml and 300ml respectively .

(b)

Mix the desired amount of potassium buffer , EDTA and NaCl solutions .

Dilute solution/mixture to 2L with water.