Instructions

Product pretreatment    

Dextran gel filtration chromatography media is supplied as a dry powder, and must be swelled in excess buffer solution before use (swell overnight at room temperature, or in boiling water for 1 hour.) Avoid magnetic stirring, or stirring with overhead stirrer or glass rod that may cause damage to the chromatography media. After swelling, adjust with buffer to form a thick slurry, about 75% (volume percentage), and degas if possible.

Column packing

Column loading should be done according to standard operating procedures. Before packing the column, it is necessary to ensure that each material is at its working temperature, and the temperature of the buffer and media must be the same (room temperature). Dextran gel filtration chromatography media comply with Darcy's law, if the flow rate is doubled, the column pressure will also be doubled.

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) The separation of sample components by the media is carried out according to the molecular size of the components, and the ones with the larger molecular size flow out first.

2) The sample feeding for gel filtration is generally 5% of the column bed in volume, and we recommend that the initial sample feeding be controlled at 1-2% of the bed volume, which can be adjusted depending on the separation situation. When desalting, the sample feeding can reach 20% of the column bed volume. The selection of column height is also related to the separation requirements. The column height should be controlled below 40 cm. Too high a gel layer will cause greater back pressure and should be avoided as much as possible.

Difficult-to-separate substances must have a certain column height and flow rate control. A 5:1 height-to-diameter ratio for desalting is advised.

3) Samples with particulates and precipitate should be filtered or centrifuged before the chromatography purification. The viscosity of the sample should not be too high, otherwise the separation efficiency will be reduced.

Elution

Elution can be done with salt-free water or with equilibration buffer as eluent, depending on the scope of the experiment. Complete separation can be achieved by adding NaCl to the equilibration buffer for gradient elution, or salt gradient elution.

Cleaning-In-Place (CIP)

After the media is used 10 times, a CIP is performed to remove the precipitated and stubborn residual proteins in the column bed. It I recommended to backwash with 4 bed volumes of 1M NaOH at 40cm/h, and then regenerate with at least 3 bed volumes of equilibration buffer.

Storage

Seplife®DX 25/100 dry powder should be stored in a dry, ventilated and clean place at 4~30℃; the hydrated media should be stored in 20% ethanol solution. Avoid contact with oxidants and 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