Recombinant Vascular Endothelial Growth Factor Agarose

Catalog Number:
LT12011-02
Packing Details:
3 ml prepacked column, or 5 ml prepacked column
Support:
6% highly cross-linked spherical agarose
Ligand:
Recombinant Vascular Endothelial Growth Factor
Ligand Density:
0.25~1.0 mg/ml
Particle Size:
45~165 μm
Maximum Flow Rate:
300 cm/h
Recommended Linear Flow Rate:
50~150 cm/h
pH Limits:
pH 2.5~9
Storage:
4 °C to 8 °C in 20% ethanol
Description:

VEGF was immobilized onto solid supports of sepharose beads for screening compounds interacting with VEGF.

Protocol of Capturing Molecules Interacting with VEGF165

  1. Wash the prepacked 5 ml column with 5~10 column volumes of distilled water to remove 20% ethanol.
  2. Equilibrate the prepacked 1 ml column with 5~10 column volumes of Binding Buffer: 20 mM sodium phosphate, 150 mM NaCl, pH 7.4.
  3. Filtrate 3-5 ml target samples through a 0.45 μm filter and load the sample.
  4. Wash with 5-10 column volumes of Binding Buffer: 20 mM sodium phosphate, 150 mM NaCl, pH 7.4.
  5. Wash with 10 column volumes of elution buffer I (100 mM sodium citrate, pH 6.0) and followed by 10 column volumes of elution buffer II (100 mM sodium glycine, pH 2.7).
  6. Discard first 1/3 column volume of elution buffer II. Immediately collect 1 column volume of eluant by elution buffer II. Neutralize collected fractions with neutralization buffer (1M Tris-HCl, pH 9.0).
  7. HSA affinity column was used as a control.
  8. Analyze the elute fraction by HPLC or other chromatography techniques to ensure that target molecule is captured.
Notes:
  1. Use the prepacked column for easier process.
  2. The target molecule should be stable under the selected elution conditions.
  3. Avoid air bubbles in the column.
  4. It is recommended to regenerate the resin by 4 column volumes of 0.1 M glycine-HCl (pH 2.7) after 5 separation cycles.
  5. 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.
  6. 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.
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