Pretreatment of Dialysis Membranes for 2D Crystallization
By Po-Lin Chiu 09/07/2012
Cut dialysis membrane tubing (Spectra/Por 12-14 kDa, Cat# 132678) into an approximate length (can be about 70 cm) and incubate them in Milli-Q water for 20 minutes.
Place in a 2-l beaker containing 50% ethanol and bring to boil.
Boil for 60 minutes. Note that a smaller beaker can be placed inside the 2-l beaker to submerge the dialysis tubing in the ethanol solution.
Pour off ethanol and rinse the tubing with 4X with Milli-Q water or until all the ethanol has been removed.
Boil the tubing in 1 mM EDTA (M.W. 292.24) and 10 mM sodium carbonate (M.W. 106) at pH 8.0 for 1 hour. In a 1.5-liter buffer, prepare 0.438 g EDTA and 0.159 g sodium carbonate.
Rinse with Milli-Q water as Step 4.
Boil the tubing in Milli-Q water for 1 hour.
Tip off the final water rinse and replace it with cold Milli-Q water containing 0.05% NaN3.
Store at 4°C refrigerator.
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