||3 ml prepacked column, or 5 ml prepacked column|
|6% highly cross-linked spherical agarose|
||Recombinant Human Fibroblast Growth Factor-Basic|
| 300 cm/h|
Recommended Linear Flow Rate:
|50~150 cm/h |
| pH 2.5~9|
| 4 °C to 8 °C in 20% ethanol|
rh-bFGF was immobilized onto solid supports of sepharose beads for screening compounds interacting with bFGF, providing an efficient and rapid technique to identify antineoplastic components from lysates, herbs or other extracts.
Protocol of Capturing Molecules Interacting with rh-bFGF
- Equilibrate the prepacked 1 ml column with 5~10 column volumes of 20 mM PBS, pH7.4.
- Filtrate 10 ml target samples through a 0.45 μm filter and load the sample.
- Wash with 5 column volumes of 20 mM PBS, pH7.4.
- Wash with 4 column volumes of 100 mM sodium glycine, pH 2.7.
- Discard first 1/3 column volume of elution buffer II and immediately collect 1 column volume of eluant and neutralize collect fractions with neutralization buffer.
- HSA affinity column was used as a control.
- Analyze the elute fraction by HPLC to ensure that target molecule is captured.
- Use the prepacked column for easier process.
- The target molecule should be stable under the selected elution conditions.
- Avoid air bubbles in the column.
- It is recommended to regenerate the resin by 4 column volumes of 0.1 M glycine-HCl (pH 2.7) after 5 separation cycles.
- After 10 separation cycles, wash the resin with 5 column volumes of 20% ethanol and 5~10 column volumes of 70% ethanol sequentially to remove hydrophobic substances.
- For longer periods of storage, keep at 4~8ºC in 20% ethanol.
|For research purposes only. Not for human or animal therapeutic or diagnostic use.|