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    Troubleshooting for Columns

    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 pessure 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

    Choose the column you are using:


Prepacked Columns