In section 24 a buffer Containing 50 mM Tris HCl 500mM NaCl

In section 2.4, a buffer Containing 50 mM Tris HCl 500mM NaCl, 250mM imidazole was wed. How would you make a 1.5 liter Solution of this same buffer Is IMAC purification the best way to go/or most efficient/best way to purify Cusf?

Solution

Answer:

6. For 1.5 L of the buffer, the calculation is as under:

Tris: MW = 121.14 gm/mol

For 1M:

1000mL of the solution requires 121.14 gm

So, for 1500mL of the solution, tris = (121.14/1000) * 1500 = 181.71 gm

For 50mM, we require tris = (181.71/1000) * 50 = 9.0855 gm

NaCl: MW = 58.44 gm/mol

For 1M:

1000mL of the solution requires 58.44 gm

So, for 1500mL of the solution, NaCl = (58.44/1000) * 1500 = 87.66 gm

For 500mM, we require NaCl = (87.66/1000) * 500 = 43.83 gm

Imidazole: MW = 68.077 gm/mol

For 1M:

1000mL of the solution requires 68.077 gm

So, for 1500mL of the solution, Imidazole = (68.077/1000) * 1500 = 102.1155 gm

For 250mM, we require Imidazole = (102.1155/1000) * 250 = 25.5288 gm