SpinDesalt Column-10
SpinDesalt Column-10 is a disposable, easy-to-operate purified product suitable for desalting, buffer replacement or small molecule removal of proteins or other large molecule samples with molecular weight greater than 5000Da.
The SpinDesalt Column-10 contains 10ml Smartdex G-25. Smartdex G-25 series resin is a kind of dextran as matrix of gel filtration chromatography resin, 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. Product use eliminates the need for chromatographs, manual loading, centrifugation operations, waiting for lengthy gravity column passes, sample collection by component, and the ability to process multiple samples in minutes.
Table 1. Characteristics of SpinDesalt Column-10
Item | Description |
Medium | Smartdex G-25 |
Average particle size D50 (μm) | 120-140 µm |
Column volume | 10 ml |
Maximum loading quantity | 2.5 ml |
Desalting efficiency | ＞90% |
Exclusion limit | Mr 5000 |
Storage | 2-8℃ |
Table 2.Composition of the SpinDesalt Column-10
Component name | Specifications (1 Piece) | Specifications (5 Piece) | Specifications (20 Piece) |
SpinDesalt Column | 1 Piece | 5 Piece | 20 Piece |
SD-10 Adapter | 1 | 2 | 4 |
2.1 Buffer Preparation
The binding buffer can be selected as the customer (e.g. PBS, etc.). It is recommended to filter the buffers by passing them through a 0.22 μm 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 Desalting column preparation
1) Open the lower plug and upper cover of the desalting column, attach the desalting column to the SD-3 Adapter, and place it in a new 15 ml centrifuge tube.
2) Centrifuge 1500×g for 1 minute and pour away the protective liquid. Mark the outer tube wall at the top of the inclined plane of the compacted resin. All subsequent centrifugation operations place the column into the centrifuge with the markings facing outward.
Note: It is normal for the resin to appear compacted or off-wall after centrifugation.
3) Add 2 ml of equilibrium solution or ultrapure water to the desalting column. centrifuge at 1500×g for 1 min to remove the equilibrium solution or ultrapure water.
4) Repeat step 3 at least three times and discard the liquid from the 15 ml centrifuge tube.
2.4 Sample desalting
Place the equilibrated desalting column from step 2.3 in a new 15 ml centrifuge tube, place it vertically, slowly add the sample dropwise to the center of the resin with a pipette gun, let it stand vertically, and when all of the sample is in the resin, centrifuge it at 1500×g for 1 min, collect the flow-through sample, and discard the used desalting column.
Questions | Cause analysis | Recommended Solutions |
Sample or equilibrium fluid does not flow through the resin | Centrifugal operating problems | Ensure that the centrifuge is working properly. |
Make sure you remove the lower plug and the top cap. | ||
Poor desalination/liquid exchange efficiency | Inaccurate samples | Samples can be added to the center of the resin ramp with the tip of the gun touching the resin. |
Avoid sample contact with the edge of the column during spiking. | ||
Incorrect centrifugal separation | For centrifuges with angular rotors, place the column markers facing outward for each centrifugation. | |
Do not exceed the recommended centrifugation speed and time. | ||
Low protein yield | Protein instability | Balance the column by replacing it with a suitable balancing fluid. |
Product | Cat. No. | Size |
SpinDesalt Column-10 | SEC03201 SEC03202 SEC03203 | 1 Piece 5 Piece 20 Piece |
SD-10 Adapter | SLM054 | 1 Piece |
