Superdex™ 75 Increase 5/150 GL
Superdex™ 75 Increase 10/300 GL, 5/150 GL, and 3.2/300 are prepacked columns for versatile use in size exclusion chromatography applications. The columns are intended for use in preparative purification as well as for characterization and analysis of proteins with molecular weights between Mr 3000 and 70 000, such as recombinant tagged proteins.
Consumables
| # | Product Name | Product Code | Price | |
|---|---|---|---|---|
| Gel Filtration LMW Calibration Kit | 28403841 | 835.37 USD |
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| Gel Filtration HMW Calibration Kit | 28403842 | 835.37 USD |
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Accessories
| # | Product Name | Product Code | Price | |
|---|---|---|---|---|
| Fingertight Connector 1/16" Male for Tubing o.d. 1/16" | 18111255 | 123.16 USD |
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| Filter Tool | 18115320 | 166.05 USD |
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| Tricorn Storage/Shipping Device | 18117643 | 139.40 USD |
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| Tricorn 10 Filter Kit | 29053612 | 194.75 USD |
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Troubleshooting
Find solutions to product related issues. For unlisted issues please contact local Cytiva service representation.
General advice to achieve good performance
Before using the column make sure that:
- Correct system has been selected in UNICORN System Control
- Correct wavelength has been set for UV/UPC monitor
- All tubing has been properly connected and tubing is not longer than needed
- All connectors are free from leakage, verified by passing a leakage test
- No tubing is folded or twisted
- Online filter, if used, is changed on a regular basis
- Correct buffers are used for the chosen columns and proteins
- All inlet tubing has been immersed in correct buffer solutions
- Enough buffer has been prepared
- Buffers have been equilibrated to the environment temperature
- Buffers/eluents have been degassed if necessary (e.g., in RPC runs)
- Suitable columns have been selected for the target proteins
- Column meets pressure requirements for selected medium
- Columns have been cleaned and prepared according to column instructions
- Samples have been clarified by centrifugation and/or filtration prior to sample loading
- Samples have been adjusted to binding buffer conditions
- Auto sampler (if used) has been prepared according to user manual
- The fraction collector has been filled with appropriate number of microtiter plates or tubes
- Appropriate arrangement for waste handling has been prepared
Superdex 75, Superdex 200 Increase and Superose 6 Increase
| Possible cause | Suggested remedy |
|---|---|
Increased back-pressure over the column and/or loss of resolution. |
Confirm that the column is the cause (see below). If so, clean it according to the procedure described in the section Cleaning in place (CIP). To confirm that the high back-pressure in the system is caused by the column, disconnect one piece of equipment at a time (starting at the fraction collector) with the pumps running. Check the pressure reading after each piece has been disconnected to determine the source of the back-pressure. |
| Possible cause | Suggested remedy |
|---|---|
Air in the column. |
Note that small amounts of air will normally not affect the performance of the column. Run 3 to 4 column volumes (CV) of well de-gassed eluent in an upflow direction at a flow rate of 0.2 ml/min (5/150) or 0.5 ml/min (10/300) at room temperature. |
| Possible cause | Suggested remedy |
|---|---|
Space between gel bed and adapter. |
Turn down the adapter to the gel bed. Perform a Column performance control. |