Prepacked Desalting Column
The Prepacked Desalting Column is suitable for desalting, buffer displacement or small molecule removal of proteins with molecular weight greater than 5000 or other large molecule samples.
The Prepacked Desalting Column contains 8.3 ml Smartdex G-25. Smartdex G-25 series medium is a kind of dextran as matrix of gel filtration chromatography medium, the main working principle is to use a mesh structure of dextran gel molecular sieves, depending on the molecular size of the separated material for separation.
The Prepacked Desalting Column selects microspheres with uniform particle size as the filling medium to make the liquid flow through fast and stable, with a single sample desalting time of about 10 min.
Table 1. Characteristics of Prepacked Desalting Column
Item | Description |
medium | Smartdex G-25 |
Average particle size D50 (μm) | 200-250 |
column volume | 8.3 ml |
Maximum loading quantity | 2.5 ml |
desalting efficiency | ＞90% |
Gravity flow rate range (pure water) | 1.5-2.5 ml/min |
Exclusion limit | Mr 5000 |
Storage | 2-8℃ |
2.1 Buffer Preparation
Water and chemicals used for buffer preparation should be of high purity. It is recommended filtering the buffers by passing them through a 0.22 or 0.45 μm filter before use.
2.2 Sample Preparation
It is recommended to filter the sample solution by passing them through a 0.22 μm or 0.45 μm filter before use.
2.3 Sample Desalination
Fix Prepacked Desalting Column and remove the bottom plug and top plug. Allow storage buffer to drain from resin by gravity flow.
Add Binding Bufffer (choose appropriate solution independently) for equilibration, repeat this step 5 times, use about 3 column volumes of Binding Buffer totally.
You can add 1.0-2.5 ml of sample to the column tube, with a maximum sample loading of 2.5 ml, and dilute the sample to 1.0-2.5 ml with Binding Buffer when the volume is insufficient.
After the sample enters the packing, continue to add the Binding Buffer for elution, collect the eluate in separate tubes, starting with the addition of the sample, collecting 2.0 ml at the front end, and then collecting the sample at a fixed volume (0.5-1 ml/tube is recommended), and test the protein concentration and salt concentration in each tube to calculate the recovery and desalting efficiency. Equilibrium solution was added at least until protein elution was complete before stopping.
3.1 Cleaning-in-place (CIP)
In order to remove residual impurities, such as denatured proteins or lipids, it is sometimes necessary to clean the desalted column in situ if the flow rate of the column becomes slower or the desalting effect becomes less effective. The commonly used cleaning solution is 0.1-0.2 M NaOH or non-ionic detergent.
1) 3 column volumes deionized water；
2) 1 column 0.1-0.2 M NaOH, 0.2 ml/min;
3) 10-15 column volumes deionized water, rinse to neutral pH.
3.2 Preservation
Rinse the medium with at least 3 column volumes of sterilization water, then cover the upper and lower plugs respectively, and store the medium in sterilization water (addition of appropriate antibacterial agents) at 2-8℃.
Product | Cat. No. | Size |
Prepacked Desalting Column | SEC003C1 SEC003C2 SEC003C3 | 8.3 ml，1 Piece 8.3 ml，5 Piece 8.3 ml，20 Piece |
