• Antigen Preparation
• General
• Antibody Usage
• Purifications
• About Pacific Immunology

Purifications

• What is the difference between an IgG purification and an affinity purification?
  • An IgG purification isolates all IgG in the serum, regardless of the specificity. This is advantageous for applications where removing serum proteins will help reduce background noise, but affinity purification against the immunizing protein isn’t possible. The affinity purification, by contrast, is advantageous for isolating only those antibodies that recognize a specific epitope. With a peptide, this can yield antibodies that have the same specificity as monoclonal antibodies.
• Is the affinity column included so that I can run additional purifications?
  • Yes, the affinity column will be shipped with the affinity-purified antibodies.
• How many times can the affinity column be used?
  • Although the capacity does decrease slightly with each use, the column can often be used for at least 10 additional purifications. We recommend storing the column at 4o C (avoid freezing or drying) and with PBS azide (added by default). In addition, it can help to filter the serum through a membrane (.45) before running it through the column (to help prevent the build up of particulates in the column). The column should also be washed between uses to remove any protein.