SpinDesalt Column-P96
SpinDesalt Column-P96 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-P96 contains 600 μl 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. The product can desalinate up to 96 samples simultaneously in a centrifugation-only operation, which is fast and high-throughput.
Table 1. Characteristics of SpinDesalt Column-P96
Item | Description |
Medium | Smartdex G-25 |
Average particle size D50 (μm) | 120-140 µm |
Column volume | ~600 μl |
Maximum loading quantity | 180 μl |
Desalting efficiency | ＞90% |
Exclusion limit | Mr 5000 |
Storage | 2-8℃ |
Table 2.Composition of the SpinDesalt Column-P96
Component name | Specifications (1 Plate) | Specifications (5 Plates) |
SpinDesalt Column-P96 | 1 Plate | 5 Plates |
96 Deep Well Plate | 2 Plates | 10 Plates |
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) 1) Open the lower plug and upper sealing film of the desalination plate, rack the desalination plate on the 96-well plate, and then fix it in the centrifuge rotor.
2) Centrifuge 1500×g for 1 minute and pour away the protective liquid.
Note: It is normal for the resin to appear compacted or off-wall after centrifugation.
3) Add 400ml 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 96-well plate.
2.4 Sample desalting
Place the equilibrated desalting column from step 2.3 in a new 96-well plate, 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 plate.
2.5 Purification Flowchart
Figure 1. SpinDesalt Column-P96 Usage Procedure
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-P96 | SEC04701 SEC04702 | 1 Plate 5 Plates |
