Instructions

Column packing

Column packing should be done according to standard operating procedures. It is important to ensure that each material is at its working temperature, , and when possible, the chromatography media may 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 exactly the same as the buffer. The equilibration solution should be a low concentration (20-50mM) buffer such as Tris or PBS.

Sample feeding

1. The sample is prepared in the equilibration buffer; turbid sample should be centrifuged and filtered before loading. Samples with high conductivity (too high salt concentration) should be processed before loading.

2. Generally, the target product is bound to the media, the impurities are washed away with the equilibration buffer solution, and then an eluent is selected and used to wash off the target product.

3. The extent to which the media adsorbs sample components depends on the charged nature of the sample, the ionic strength and pH of the mobile phase. The lower the salt concentration, the stronger the adsorption of the sample components by the media.

Elution

Elution can be carried out by increasing the salt concentration or changing the pH value. The method of increasing the salt concentration is often used for elution.

Regeneration

Generally, use high salt concentration buffer (containing 1~2mol/L NaCl) or lower the pH to wash more than 10 times the volume of the column. Then wash with the equilibration solution that was used for binding proteins until the equilibrium is reached.

If there are inactivated proteins or lipids that cannot be washed away during regeneration, they 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, first wash with 1 BV of 0.1M NaOH, and then wash with equilibrium buffer solution until the pH is neutral.

3. For proteins and lipids with strong hydrophobic binding, wash with 4-10 BV of 70% ethanol or 30% isopropanol. It should be noted that the concentration of the organic solvent should gradually increase to avoid bubbles.

Storage

Sealed and stored at 2-8°C (preservation solution is 20% ethanol) in a ventilated, dry and clean place, do not freeze.

Ordering information

Production date: See label

Expiry Date : 5 years, under proper storage conditions

Manufacturer: Sunresin New Materials Co. Ltd.

Add:No. 135, Jinye Rd, Xi'an Hi-tech Industrial Development Zone, Shaanxi, 710076, China

www.seplite.com     www.sunresin.com

E-mail: info.lifescience@sunresin.com