Focurose 30PG

Resin

Matrix Highly cross-linked agarose and dextran

Particle size range 

25-45µm

Average particle size (D50)

35±5µm

Separation range (globular proteins)

≤10kDa

pH stability

3-12 (long-term) 1-14 (short-term)

Chemical stability

All common solutions: 8M urea, 6M HCl-guanidine, 70% ethanol, 1M NaOH

Storage solution

0.2M NaAc 20% ethanol

Storage conditions

4℃-30℃

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