Bio-Rad Profinity eXact™ Purification and Tag Cleavage Consumables - Manual (Page 16)

Problem Possible Cause Solution
Precipation during Binding capacity of Load less sample
purification cartridge exceeded
Protein aggregation Include detergent or
glycerol in buffers
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Section 11
Troubleshooting Guide
Problem Possible Cause Solution
Cartridge clogging Particulates in samples Filter all samples and buffers
or slow flow rate or buffers through 0.45 µM filter prior
to application
Sample too viscous Add nuclease to lysate to
degrade DNA
No target protein Low level of target Check expression level by
in eluant protein in starting SDS-PAGE
material
Target protein not Used resin must be
binding regenerated before reuse.
Follow regeneration protocol
in Section 10
Inaccessible tag Clone into pPAL’s Spe I site
Target protein in Intrinsic cleavage Ensure no chloride ions are
flowthrough or wash present in lysate or bind/wash
fractions buffers. Perform purification
at 4°C. Clone into pPAL’s
Spe I site
Uncomplete elution No or slow cleavage Lengthen cleavage incubation
step. Clone into pPAL’s Spe I
site
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