Neozene offers custom polyclonal antibodies to customers locally and globally, including researchers at universities and in biotechnology and pharmaceutical companies. Listed below are the different custom antibody production services we offer. Complete confidentiality of all orders is assured.
Polyclonal Antibody Production for 2 rabbits:
• NIH 90-day standard protocol or 70-day conventional protocol
• 10 mL pre-immune serum
• 100 mL anti-serum

Additional Services:
• ELISA
• Peptide Synthesis10 mg peptide, >80% purity, MS and HPLC certified
• Peptide-Carrier Protein Conjugation to BSA or KLH

Complete Anti-Peptide Antibody Package includes:
• Free consultation for selection of epitope
• 5 mg purified (>80% purity, up to 15 amino acids) peptide delivered to researcher for immediate study
• 1 mg conjugated to BSA for ELISA screening; 4 mg conjugated to KLH for antibody production
• Immunization of rabbits, mice, or chickens at your preference
• Peptide-specific ELISA titration of antibody for all bleeds
• Pre-immune samples of serum, ascites, or eggs
• Shipment of 100 mL anti-serum, 20 mL ascites, or 12 antibody-containing eggs

Note: The rabbits can be maintained on a 3-4 wks cycle of immunization and bleed as required by the project. The results of your testing will determine whether the project should be continued, modified, or terminated.

Protocol

Subject:	New Zealand White Rabbits
Adjuvant:	Complete Freund's Adjuvant (CFA) initially, followed by incomplete Freund's Adjuvant (IFA) for all subsequent injections.
Immunogen:	Typically, proteins and conjugated peptides. Amount of immunogen used for each immunization is 50 to 500 μg.
Procedure:	The immunogen is diluted to one mL with sterile saline and combined with one mL of the appropriate adjuvant. The antigen and adjuvant are mixed thoroughly to form a stable emulsion which is injected subcutaneously and provides enhanced immune response from the sustained presence of the immunogen.
Bleeds:         Schedule:	Blood is collected from the central ear artery with a 19 gauge needle and allowed to clot and retract at 37°C overnight. The clotted blood is then refrigerated for 24 hours before the serum is decanted and clarified by centrifugation at 2500 rpm for 20 minutes. Time 0 Bleed 25 mL (yields 10 mL pre-immune serum).                         Immunize with antigen in CFA. Week 3            Immunize with antigen in IFA. Week 6            Immunize with antigen in IFA. Week 7            Bleed 50 mL (yields 20 mL serum). Week 10          Immunize with antigen in IFA. Week 11          Bleed 50 mL.

Phosphospecific Antibody Production

Neozene offers phosphospecific antibody production which includes:
• 15 mg purified (>90%, up to 15 amino acids) phosphopeptide synthesis -- 5 mg used for
phosphospecific epitope affinity purification, 4 mg for KLH conjugation used for antibody production, 1 mg for BSA conjugation, and 5 mg delivered to researcher
• 11 mg purified (>80%, up to 15 amino acids) non-phosphopeptide synthesis -- 5 mg used for affinity depletion, 1 mg for BSA conjugation, and 5 mg delivered to researcher
• Immunization of 2 rabbits
• Phosphopeptide and non-phosphopeptide ELISA titration of antibody for all bleeds
• Pre-immune sample of serum (5 mL/each rabbit) and anti-serum (100 mL total)
• Affinity purification of selected batch of serum
• Depletion of nonphospecific peptide matrix
• Shipment of purified phosphospecific antibody which does not cross react with non-phosphopeptides (confirmed by ELISA)

Peptide-Carrier Protein Conjugation

Neozene provides the following peptide-carrier protein conjugation:
• Peptide-KLH conjugation
• Peptide-BSA conjugation
• Peptide-GGG conjugation
*Average coupling efficiency between 80-95% (HPLC analysis).