|Resin code||Seplife® 6FF|
|Appearance||White Sphere beads|
|Matrix||6% cross-linked agarose|
|pH stability||2~12 (long term), 2~14 (Short term, CIP)|
|Application||Proteins, nucleic acids and peptides downstream of |
biopharmaceuticals and bioengineering
|Chemical stability||Stable in water buffer solution: 2M NaOH; 70% ethanol, 30% |
isopropanol, 30% acetonitrile, 1% SDS, 6M guanidine
hydrochloride, 8M urea.
|Exclusion limit||10000~4×106 globulin|
|Test conditions: Chromatography column 16mm * 300mm; Column bed height 15 cm; temperature 25℃; |
mobile phase was 0.1 mol / L NaCl.
Packing is performed in accordance with standard operating procedures. It must be ensured that each material is at operating temperature and that the gel needs to be degassed before it is packed.
Equilibrate the column with 2 to 5 column volumes of the loading balance, making sure that the conductivity and pH of the effluent are exactly the same as the conductivity and pH of the loading buffer.
(1)The sample is prepared with a buffer, and the turbid sample is centrifuged and filtered for loading. Samples with excessive salt content and too small concentration should be treated first and then loaded.
(2)The separation of the sample components by the medium is carried out according to the molecular weight of the components, and the molecular weight is first flowed out.
(3)The loading volume is about 1-2% of the column volume, and the smaller the better separation performance.
Elution was carried out with buffer, and the flow rate and buffer composition were kept constant during the elution.
It is usually washed with a buffer to balance and can be used again. Some inactivated proteins or lipids cannot be washed off during regeneration and can be removed by in-place cleaning (CIP).
(1)For an ion-bonded protein, it can be removed with 0.5 to 1 column bed volume of 2 M NaCl.
(2)For precipitated proteins, hydrophobically bound proteins or lipids, it can be removed with 1 column bed volume of 0.1 M NaOH. (3) For strongly hydrophobically bound proteins, lipids, etc., it can be washed with 4 to 10 column volumes of 70% ethanol or 30% isopropanol. However, it should be noted that the concentration of the organic solvent is gradually increased in a gradient manner, otherwise bubbles are easily generated.
After the cleaning is completed, equilibrate the column with at least 3 bed volumes of equilibration buffer until the pH and conductance remain unchanged.
Sealed and stored at 4~30℃(20% ethanol in storage solution), dry, ventilated, clean, not frozen; used columns are stored in 4℃, 20% ethanol solution.
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