Dialysis after antibody purification with Bart 2-28-12

2015-01-13

azim58 - Dialysis after antibody purification with Bart 2-28-12

Dialyze sample in Slide-A-Lyzer into 2 L 0.75 M NaCl in 10 mM sodium

phosphate pH about 7.4 for 1 hr or several hours (use a 4 L beaker)
o the 10 mM sodium phosphate can be made from a 200 mM sodium
phosphate stock
o 200 mM sodium phosphate stock can be prepared by diluting 28.39 g
Na2HPO4 into 1 L. This can be adjusted to pH 7 (the pH may gradually
increase over time as the solution ages) with 1 M HCl (required about
10 mL of this last time I adjusted)

Add 1 L 10 mM sodium phosphate to the dialysis beaker (now 0.5 M

NaCl) for 1 hr or overnight

dialyze into 0.25 M phosphate
dialyze into 0.125 M phosphate
dialyze into 10 mM phosphate or just PBS
Concentrate using BSA if necessary
dialyze into 10 mM phosphate or just PBS