Product Introduction
Focurose 75PG is a size exclusion resin suitable for the fine purification of biomolecules in both laboratory and industrial-production scale.
Features
Focurose 75PG Performance Parameters
Resin
|
Highly
cross-linked agarose and dextran |
Particle
size range |
25-45
µm |
Average
particle size (D50) |
35±5
µm |
Separation
Range (Globular Proteins) |
3-70kDa |
pH
stability |
3-12
(long-term) 1-14 (short-term) |
Chemical
stability |
Compatible
with all common solutions including 8M urea, 6M guanidine hydrochloride, 70%
ethanol, and 1M sodium hydroxide |
Storage
solution |
0.2M
NaAC, 20% ethanol |
Storage
conditions |
4℃-30℃ |
Frequently Asked Questions and Solutions
Issue |
Possible Causes |
Solutions |
Resolution
difference between target peak and impurity peak |
Overloading of sample |
Reduce the
sample volume to 0.5% of the column volume (CV) |
Sample too viscous |
Dilute the sample
appropriately |
|
Excessive flow rate |
Reduce the flow
rate |
|
Short column length |
Choose a longer
or narrower column |
|
Excessive dead volume |
Minimize dead
volume in tubing and connectors |
|
Poor column packing |
Repack the
column or use prepacked columns |
|
Unfiltered sample |
Filter the
sample using a 0.22 µm or 0.45 µm filter |
|
Contaminated resin |
Clean the column
and re-equilibrate |
|
Improper column installation |
Reinstall the
column vertically |
|
Non-uniform temperature usage |
Maintain a
constant temperature throughout the process |
|
Unexpected
elution peak |
Inconsistent sample volume |
Maintain the
same sample volume throughout |
Ionic interaction between protein and resin |
Maintain the
ionic strength of the buffer in the range of 0.05-0.15 NaCl |
|
Ionic interaction between protein and resin |
Reducing the
ionic strength can minimize hydrophobic interactions. Additionally,
hydrophobic interactions can be reduced by increasing the pH, adding
surfactants, or incorporating organic reagents |
|
Changes in the sample during storage |
Prepare fresh
samples |
|
Precipitation of proteins and lipids in the chromatography column |
Clean the column
or replace it with a new one |
|
Microbial growth in the resin |
The column does
not support microbial growth during usage, but it should be stored in 20%
ethanol |
|
Washing peak
elution occurs earlier |
Gaps in the packed column |
Repack the
column |
Formation of protein dimers or oligomers |
Ensure the
stability of the sample under experimental conditions |
|
Delayed elution
of the washing peak |
Ionic or hydrophobic interactions between proteins and the resin |
Maintain an
ionic strength of 0.05-0.15M NaCl in the buffer |
Dirt on the resin, filter membrane, or top of the column bed |
Clean the column
and re-equilibrate |
|
Microbial growth in the resin |
The column does
not support microbial growth during usage, but it should be stored in 20%
ethanol |
|
Rapid increase
in chromatographic peak |
Overly tight packing of the resin |
Repack the
column |
Slow or tailing
chromatographic peak |
Loose packing of the resin |
Repack the
column |
Cracks or
dryness in the column bed |
Leakage or introduction of large air bubbles |
Check for leaks
or bubbles in the tubing and repack the column if necessary |
Slow liquid flow |
Aggregation of proteins or lipids |
Clean the resin
or membrane promptly |
Protein precipitation in the resin |
Adjust the
composition of the equilibration and elution buffers to maintain the
stability of the target substance and the binding efficiency of the resin |
|
Microbial growth in the separation column |
All reagents
used must be filtered and degassed. The sample must be centrifuged or
filtered before applying it to the column |
|
Compression of the column bed |
Repack the
column |
|
Column bed
contains bubbles |
Repack the column due to temperature differences during use or
residue in the tubing |
Repack the
column |
Increased
pressure |
Sample turbidity |
Prepare a fresh
sample |
Clogging in the tubing or frit |
Clean the
tubing, frit, and resin; repack the column |