Custom Polyclonal Antibody Services

Polyclonal Antibody Service

High quality, highly specific antibodies

AMSBIO is a leading manufacturer of off the shelf and custom antibodies. We have completed thousands of projects for industry and academia in the last 20 years.

Our polyclonal antibody services provide a fast and economical solution with extensive flexibility to incorporate desired properties. You can provide your own immunogen or work with our world-class synthesis peptide chemistry group to design an epitope-specific and immunogenic peptide to meet your specifications.

AMSBIO can also develop a pre-optimized antibody validated in the assay of your choosing.

Our Services

Deliverables

              • Peptide and phospho peptide 2-3mg each
              • 0.5-1ml of pre-immune sera and antisera per rabbit
              • Project report with ELISA results and dot blot
              • Non-phospho antibody 3-5ml
              • 1-5mg of affinity purified phospho-specific antibody

Service includes

              • Immunization of 2 New Zealand rabbits
              • 2x (0.5-1ml) pre-immunization samples
              • 2x (0.5-1ml) post-immunization samples
              • Synthesis of peptides and KLH conjugation

Adjuvant: Complete Freund’s Adjuvant (CFA) initially, followed by Incomplete Freund’s Adjuvant (IFA) for all subsequent injections.

Immunogen - KLH conjugated phospho-peptide. 250µg immunogen is used for each immunization.

Procedure

The immunogen is diluted to 0.5 ml with sterile saline and combined with 0.5 ml of the appropriate adjuvant.

The antigen and adjuvant are mixed thoroughly to form a stable emulsion that is injected beneath the skin of the rabbit (subcutaneously) in the area around the shoulders and intra-muscularly into the large muscle of the rear legs (that is about 1/4 of the antigen being used in each area).

This provides enhanced immune response from the sustained presentation of the immunogen.

Bleeds

Blood is collected from the central ear artery with a 19-gauge needle and allowed to clot and retract at 37°C for 1h followed by 12h at 4°C. The clotted blood is then decanted and clarified by centrifugation at 10,000 RPM for 10 minutes.

Schedule

Week 0 : Bleed 1-2 ml (yields 0.5-1 ml pre-immune serum) Immunize with 250-350mg/rabbit antigen in CFA.

Week 2 : Immunize with 250-350 mg antigen / rabbit in CFA.

Week 4 : Immunize with 100-150 mg antigen / rabbit in IFA.

Week 5 : Immunize with 100-150 mg antigen / rabbit in IFA.

Week 6 : Test bleed 0.5-1 ml. If ELISA data OD value, 1:4000 lower than 2.0,  1:6000 lower than 1.0, re-immunize antigen / rabbit in CFA.

Week 7 : 1st Production bleed 3-5 ml. Immunize with 100-150 mg antigen / rabbit in IFA. ELISA test again if previous tests were negative. The continuation of the project needs to be discussed if negative at this stage.

Week 8 : 2nd Production bleed 3-5 ml. Immunize with 100-150 mg antigen / rabbit in 0.9% NaCl-H2O.

Week 9 : 3rd Production bleed 3-5 ml. Immunize with 100-150 mg antigen / rabbit in 0.9% NaCl-H2O.

Week 10 : Terminating bleed 40-60 ml/rabbit. A total of 60-80ml/rabbit is collected.

Sera is then shipped to customer or purified either using Protein A/G or Ammonium Sulphate. Peptide affinity purification can be added at this stage.

Notes

The rabbits can be maintained on a cycle of immunizations and bleeds as required by the project. The results of testing will determine whether the program should be continued, modified, or terminated. Extra bleeds can be taken upon customer’s requirement of extended project with extra charge.

Protocols are subject to change.

Peptide & Antibody Request Form

Let's get started

Complete our service request form and we will contact you to discuss your requirements.