Dextran Desalting Prepacked Column G25
Dextran Desalting Prepacked Column G25 is used for desalting proteins with molecular weight greater than 5000 or other large molecule samples, replacement of buffer solution, or removal of small molecules.
Dextran Desalting Prepacked Column G25 is packed with 1ml or 5ml of 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.
Table 1. Characteristics of Dextran Desalting Prepacked Column G25
|
Item |
Description |
|
Medium |
αdex G-25 |
|
Average particle size D50 (μm) |
120-140 μm |
|
Maximum loading quantity |
10% column volume |
|
Desalting efficiency |
＞95% |
|
Exclusion limit |
Mr 5000 |
|
Storage |
2-8℃ |
2.1 Buffer Preparation
Water and chemicals used for buffer preparation should be high purity. It is recommended to filter the buffers by passing them through a 0.22 μm or 0.45 μm filter before use.
The binding buffer is the solution used to protect the protein during the desalting. Choose as customer needs.
2.2 Sample Preparation
Water and chemicals used for buffer preparation should be high purity. It is recommended filtering the buffers by passing them through a 0.22 μm or 0.45 μm filter before use.
2.3 Sample Desalting
The Dextran Desalting Prepacked Column G25 is a pre-packed desalting column. Various conventional chromatography systems can be used to introduce the use of the Dextran Desalting Prepacked Column G25 method by taking KTA instrument as an example.
1) Fill the syringe or pump tubing with binding buffer. Remove the stopper and connect the column to the syringe (with the provided connector), or pump tubing, “drop to drop” to avoid introducing air into the column. Remove the snap-off end at the column outlet.
2) Wash the column with at least 5 column volumes of binding buffer at 0.2 ml/min or 1 ml/min for 1 ml and 5 ml column respectively.
3) Apply 10% column volume of sample, using a syringe fitted to the connector or by pumping it onto the column at the same flow velocity. Dilute it with binding buffer to the corresponding volume if the sample volume is not enough.
4) Elute with binding buffer and collect eluent by fixed volume. Test the Sample recovery and desalting efficiency.
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 0.1-0.2m NaOH or non-ionic detergent.
1) 3 column volumes deionized water；
2) 1 column 0.1-0.2 M NaOH,0.2ml/min;
3) 10-15 column volumes deionized water,rinse to neutral pH.
Note：The viscosity of 0.1–0.2 M NaOH may increases the column pressure. Customers can reverse flushing according to need.
3.2 Preservation
Rinse the medium with at least 3 column volumes of 20% ethanol solution, then cover the upper and lower plugs respectively, and store the medium in 20% ethanol solution at 2-8℃.
|
Product |
Cat. No. |
Size |
|
Dextran Desalting Prepacked Column G25 |
SEC018C11 SEC018C51 SEC018C15 SEC018C55 SEC018CS |
1×1 ml 5×1 ml 1×5 ml 5×5 ml 3×1 ml+1×5 ml |
