Pierce Biotechnology
PO Box 117
(815) 968-0747
thermofisher.com
3747 N. Meridian Road
Rockford, lL 61105 USA
(815) 968-7316 fax
3
Troubleshooting
Problem
Possible Cause
Solution
Sample or buffer does not
flow through resin
Centrifugation problem
Ensure that centrifuge is in proper working condition
Ensure bottom closure is removed
Ensure top cap is loosened
Sample contamination
Improper sample loading
Apply sample directly to center of the resin bed; touch tip
to resin to expel all sample
Avoid contact with sides of column
Improper centrifugation
For fixed-angle rotors, place column in the same orientation
each time and do not exceed recommended centrifuge speed
Do not exceed recommended centrifugation speed or time
Low yield
Sample was not completely in
solution before adding to the
column
Centrifuge sample at 14,000
×
g
for 10 minutes before
adding to the column
Portion of protein still
remaining in spin column
Use a stacker to recover more protein (for most samples,
the majority of protein is recovered without a stacker)
Protein precipitated in
equilibration buffer
Check for protein solubility in the final buffer or solution
Protein bound to resin matrix
Use alternative resin, such as Polyacrylamide Spin
Desalting Columns (see Related Thermo Scientific Products
section)
Recovered protein or
sample is dilute
Wash/equilibration buffer was
not adequately removed
Before adding the sample make sure the wash/equilibration
buffer was adequately removed by centrifugation (i.e.,
column appears uniformly white with no solvent streaks)
Related Thermo Scientific Products
89849
Pierce
Polyacrylamide Spin Desalting Columns, 7K MWCO, 0.7mL, 25 columns
89862
Pierce
Polyacrylamide Spin Desalting Columns, 7K MWCO, 0.7mL, 50 columns
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