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ConductScience makes polyclonal antibody production easier than ever

Background information

  • Animals : New Zealand rabbits
  • Adjuvant : First injection with complete Freund’s adjuvant, then incomplete Freund’s adjuvant.
  • Immunogen : Protein or KLH-coupled peptide; each injection : 100 to 200 μg of immunogen

Methods

Immunisations : immunogen diluted with saline buffer, and mixed with the appropriate adjuvant (mass ratio is1:1). The emulsion is injected to rabbits by subcutaneous injection around the shoulders and intramuscular injection back thighs. Each injection takes about 1/4 immunogen. Then immunogen can be persistent so as to enhance the immune response

Blood : rabbit ear artery for blood serum, precipitation at room temperature overnight

Process

Day 00 : Serum collected 2ml (0.5-1ml pre-immune serum)
Day 01 : 1st -immunization, complete Freund’s adjuvant 200μg antigen
Day 15 : 2nd -immunization, complete Freund’s adjuvant 200μg antigen
Day 29 : 3rd -immunization, incomplete Freund’s adjuvant 100μg antigen
Day 35 : Serum collected 2-3ml , ELISA and WB test
Day 36 : 4th -immunization, incomplete Freund’s adjuvant 100μg antigen

  • Day 43 : 5th -immunization, incomplete Freund’s adjuvant 100μg antigen. If Elisa test is negative, 2-3ml bleeding for ELISA and WB test
  • Day 50 : 6th -immunization, incomplete Freund’s adjuvant 100μg antigen. If test is still negative, we need to discuss about project
  • Day 57 : 7th -immunization, incomplete Freund’s adjuvant 100μg antigen
  • Day 64 : last bleeding
  • Day 65 : serum purification, final tests
  • Day 69 : Delivery