Question

BUFFERS:

(Final concentration for 1x is 20 mM Tris; 500 mM NaCl; 1 mM CaCl₂; 1 mM MgCl₂; pH 7.4)

5x ConA Buffer, (1L)

• 700 ml dH₂O, set stirring and add:

• 100 ml 1M Tris, pH 7.2-7.4

• 5 ml 1M MgCl₂

• 5 ml 1M CaCl₂

• 146 g NaCl

• Check pH is between 7.2 and 7.4

• To 1000 ml with dH₂O

Elution Buffer A: ~0.2M AMM: 0.1 g alpha methyl mannopyranoside in 2 mls ConA buffer

Elution Buffer B: ~0.5M AMM: 0.2 g alpha methyl mannopyranoside in 2 mls ConA buffer

Elution Buffer C: ~1 M AMM: Add 0.4 g alpha methyl mannopyranoside in 2 mls ConA buffer (you may need more of this)

Formula weight for alpha methyl mannopyranoside is 194.2 g.

Questions:

Buffers: Calculate the concentrations of Tris, CaCl2 and MgCl2 in the 5x ConA Buffer stock

Tris:

CaCl2:

MgCl2:

For Buffer B: What is the exact concentration of the AMM to three significant figures