Jul 17, 2020
Version 2
Dialysis using D-Tubes V.2
- 1University of Manitoba
Protocol Citation: Neilier Junior 2020. Dialysis using D-Tubes. protocols.io https://dx.doi.org/10.17504/protocols.io.bhqkj5uw
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: June 20, 2020
Last Modified: July 17, 2020
Protocol Integer ID: 38380
Keywords: Dialysis, Protein Purification, Proteomics, Sample Preparation, Desalination, Purification,
Abstract
Sample preparation is crucial for successful biomolecule analysis. For this, the method must be individually chosen, based on the characteristics of the sample and the analyte. How some strategies can compromise the integrity of the molecule of interest. The dialysis D-tubes allow the concentration of samples with no precipitation by organic solvents or salts.
Materials
D-Tubes Dialyzers
Milli-Q Water
1000 mL Beaker
Automatic micropipette
Floating rack
Plastic film
Safety warnings
Wear personal protective equipment: gloves, lab coat and mask.
Before start
Organize your workspace
Make sure all solutions and equipment are available. Plan the experiment!
Material preparation
Material preparation
Choose the cutting mass (3.5 to 14 kDa) of the dialysis D-Tube based on the biomolecule to be purified.
Procedure
Procedure
Complete the dialysis D-Tube with Milli-Q water and let it equilibrate for 15 min
Remove the water and weigh the dialysis tube
Add the sample with the aid of a pipette and weigh the tube again
Prepare a beaker with 1000 mL of Milli-Q water (or exchange buffer) and a magnetic stir bar
Place the dialysis tube on a floating rack and immerse the tube in the water in the beaker
Cover the beaker with plastic film, start stirring and monitor the conductivity of the solution in the beaker. When the conductivity stops increasing, the concentrations between the sample and the solution are in equilibrium
If conductivity is not controlled: 1 h of dialysis suffices to desalinate 3 M of ammonium sulfate in these conditions
Remove the dialysis tubes from the beaker and weigh the contents