Seplife® 4AG/90 Size Exclusion Chromatography Agarose Resin

Size Exclusion Chromatography - Agarose

Appearance: White spherical beads
Type: Size exclusion agarose
Matrix: 4% crossed linked agarose
Particle size range (μm): 45-165
Selectivity: Kav IgG 150000MW: 0.66-0.76
pH stability: 3-13 (operational), 2-14 (CIP)
Chemical Stability: Stable in all common aqueous buffers; 1M sodium hydroxide; 8M urea; 6M guanidine hydrochloride; 70% ethanol.
Flow rate* (cm/h): max 420cm/h
Shipped as: 20% ethanol slurry

*Testing conditions: Chromatography column 16mm×400mm; column bed height 25cm; temperature 25°C; mobile phase water.

Column packing
Column loading should be performed in accordance with standard operating procedures. It is important to ensure that each material is at its working temperature, and the media may need to be degassed before column packing.

Equilibration
Equilibrate the column with an appropriate 2-5 column volume buffer. Ensure the conductivity and pH of the effluent are the same as the equilibration buffer.

Sample feeding
1. Samples are prepared in buffer, and cloudy samples need to be centrifuged and filtered before loading. Samples with too high salt content and too low concentration should be processed first before loading.
2.The separation of components in the sample by the media is carried out according to the molecular size of the components. The ones with the larger molecular size flow out first.
3. The sample loading volume is about 1-2% of the column volume. The smaller the loading volume, the better the separation.

Elution
Elute with buffer, keep the flow rate and buffer composition unchanged during elution.

Regeneration
Generally, wash with buffer solution to balance and the media can be used again. Inactivated proteins or lipids that cannot be washed out during regeneration, can be removed by Cleaning-In-Place (CIP).

Cleaning-in-place (CIP)
1. For proteins bound by ionic bonds, 0.5~1 BV of 2M NaCl can be used to remove them.
2. For precipitated proteins, hydrophobically bound proteins or lipids, wash with 1 BV of 0.1M NaOH.
3. For proteins and lipids with strong hydrophobic binding, wash with 4-10 BV of 70% ethanol or 30% isopropanol.
4. The concentration of the organic solvent should gradually increase to avoid bubbles.
After cleaning, equilibrate the column with equilibration buffer solution at least 3 times the volume of the column bed until the pH and conductivity remain unchanged.

Storage
Sealed and stored at 4~30°C (preservation solution is 20% ethanol), in a ventilated, dry and clean place, do not freeze.