Antibody Production FAQ > General

General

  • What is Sodium Azide and should I add it to the antiserum?
    • Sodium Azide is a preservative that prevents bacteria from growing in the serum (bacteria can produce proteases that denature any proteins in the serum, including antibodies). We typically recommend adding azide unless the serum is going to be used directly in cell culture applications. Azide can be removed by using a dialysis membrane with a molecular weight cutoff between 12,000 and 14,000.
  • How will the serum be shipped?
    • Serum is typically shipped after each bleed is taken. The serum will be frozen, spun down and shipped via FedEx overnight on ice packs.
  • What concentration of antibody can be expected?
    • In general, there will be approximately 5-10mg of IgG in each ml of serum. Of this, there will be approximately 100ug of peptide-specific antibody. So, a typical affinity purification of 25ml of serum will yield approximately 2-3mg of specific antibody.
  • Will the titer of antibody increase with additional immunizations?
    • For rabbit projects, the titers remain relatively level after the 1st production bleed and additional immunizations are used to maintain antibody titers rather than to increase them. Specificity does increase between the 1st and 3rd bleeds as part of the affinity maturation process and so antibodies from the third and later bleeds represent the most specific antibodies (which is why the affinity purification is run on the third bleed).
  • Are accelerated protocols available?
    • No, accelerated protocols are largely a marketing gimmick that overlooks the natural timeline needed for the animal’s immune system to develop high affinity antibodies.
  • What kind of rabbits / chickens does Pacific Immunology use?
    • New Zealand White Rabbits and Rhode Island Red Chickens
  • Will I receive any leftover peptide at the end of the project?
    • Yes, you will receive all leftover peptide at the end of your project, helping ensure the confidentiality of your research. Typically we synthesize 40-50mg of peptide, so you should receive 30-45mg of peptide at the end.