Fee:  $1600

     We generate polyclonal antibodies in rabbits, the best immune responders, by immunization with peptide or protein antigens, and/or cDNA-coated microparticles.  Two rabbits are injected for each antigen.  To produce a high immune response, we inject microparticles carrying the cDNA, protein or peptide antigen directly into the spleens of rabbit hosts.  The animals are boosted six or seven times every two weeks with the protein or peptide.  For peptide or protein antigen, at least 4mgs of antigen is required.  For cDNA, at least 6mgs of antigen is required. 

    The antibodies that we generate will be reactive in ELISA assays.  The cost of production will vary within the range given above depending on per-diem charges for housing rabbits in the Animal Resource Center.  Turnaround time is 4-6 months.

Polyclonal Antibody Performance:

• Polyclonal sera must elicit a positive ELISA response to the stated peptide or protein before they are considered acceptable for delivery.  A positive response is an ELISA reading with a three-fold higher reading over background at the 1:10,000 antibody titer (using 0.5mg of antigen per well). 
• Polyclonal sera are likely (but not guaranteed) to elicit a positive response by Western Blot assay of overexpressed protein as conducted by the client. We do not perform western blot assays in our facility. We ask that the results from any other assay performed by the client be sent to The Antibody Core Facility for our record keeping.
• Animals for polyclonal antibody production will be maintained for 17 weeks, during which 7 immunizations will be performed.  At such time, if the animal sera elicits an ELISA response as described above, the animal will be sacrificed. 
• Usually, three additional boosts can be scheduled if a higher antibody titer is required.  The client will be charged for the additional time required for boosting. If the client wishes to keep the animals beyond our standard protocol of 3 boosts, this must first be discussed with Dr. Lai.   
• The client will be billed for animal boarding costs of all immunized animals until the date of sacrifice. 
• The Antibody Core Facility will provide approximately 160 to 200 ml of immune serum derived from immunized animals.
• For more information, please review our Antibody Production Guidelines, speak with Dr. Lai or the Antibody Core Facility staff.

How to proceed:

1. Fill out the order form, which may be downloaded as an Adobe PDF file here. The order form must be accompanied by an IDR, together with required reagents (see below), to Dr. Wayne Lai (K3.118 or Y4.312D or FAX 214-648-2951).
2. Investigators must file an "application for use of animal subjects" with the Institutional Animal Care and Use Committee (IACUC).  Antibodies may be created using the Antibody Core's protocol.  A short form of the application may be downloaded at this site.  Investigators should consult with Dr. Lai if they require guidance in filling out and submitting this form
3. Investigators who wish to raise antibodies against a protein or peptide antigen, may do so by providing us with the NCBI or Swiss-PROT "Accession Number" for the protein, or its amino acid sequence. We will design two to three optimal peptide sequences according to the criteria of potential antigenicity, hydrophilicity and surface probability.  You may arrange to have the peptides synthesized by the Protein Chemistry Technology Core or elsewhere.  The costs of peptide synthesis is the responsibility of the investigator.
4. Order~20 mg of each peptide at standard grade (70% pure, desalted).
5. For co-immunization with cDNA, the investigator should provide 6 mg cDNA encoding the antigen cloned in PcDNA3.1 (or 1.0), which has a CMV promotor.  cDNA should be prepared by using Qiagen endotoxin-free kit and dissolved in sterile water or PBS (2µg/ul).