advantages and disadvantages of gel filtration chromatography 7 | Page 5

4. Elution later than expected
Reason: Digestion by proteases.
Recommended Solution: Add protease inhibitors to buffer.
Reason: Hydrophobic adsorption.
Recommended Solution: Add 10-20% ethanol or other organic solvents to
buffer or decrease salt concentration in high ionic strength buffer.
Reason: Ionic adsorption due to negatively charged groups on the support
(Carboxyl-, Sulphate-) and positively charged roups on sample molecules.
Recommended Solution: Include 150mM NaCl (or other salt) in buffer.
TIP: Be sure that your sample is stable in the buffer additive prior to use.
Some additives recommended here may denature or otherwise inactivate
a protein.
TIP: Be sure your column and system are stable in the buffer additive prior
to use.
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