Custom Polyclonal Antibody Production

Polyclonal antibodies are one of the most widely used research reagents. Custom polyclonal antibody production can be used for a range of research applications, including:

• western blotting
• immunohistochemistry
• immunocytochemistry
• flow cytometry
• immunoprecipitation
• neutralization assays
• ELISAs

Who we are:

Having produced more than 10,000 custom polyclonal antibodies, ProSci Incorporated has the experience and knowledgeable staff to help you develop the antibody your research needs. We produce product custom polyclonal antibodies at our animal facility less than 20 miles from our laboratory location, all located within San Diego County, California. Animal welfare, quality, and project confidentiality are assured due to the centralization of all procedures to ProSci-operated facilities.

We offer custom polyclonal antibody production packages that cover all scales of projects, from a single project for a researcher targeting one protein, to the long-term large scale production needed for diagnostics. Animals are placed on an initial immunization protocol and at the end of the period you have a three day screening period to evaluate the final scheduled bleeds. Animals can then be terminated at no additional cost, exsanguinated to harvest additional serum (~60mL per rabbit; $50), or placed on an extension schedule for longer-term production ($75 per month per animal for rabbit and chicken, $200 per month per animal for goat). Animals can be maintained on extension schedules for multiple years, allowing you to obtain as much anti-serum as is needed for your research.

ProSci can produce a custom polyclonal antibody using an antigen you provide (see requirements below), or help you to design a guaranteed peptide antigen that we can synthesize. We offer a variety of packages, including Peptide Cocktail Antibody Production and Phospho-Specific Antibody Production packages. We offer a wide range of antibody production services in three animal species:

• Rabbit
• Chicken
• Goat

Antigen Requirements:

Client-Provided Antigen Requirements: For client-provided antigens, we accept recombinant proteins, fusion proteins, ESTs (Expressed Sequence Tags), peptides (conjugated or unconjugated), enzymes, heat-killed bacteria and viruses, and cells. We can perform a complete custom polyclonal antibody production protocol with the following amounts of antigen: 0.5 mg for 2 chickens, 1.0 mg for 2 rabbits, and 1.5 mg for goat. We can start a custom polyclonal antibody production project with 50% of the total amount of antigen needed. Additional quantities can be sent during the project’s progress. If providing a fusion protein made using a large fusion partner (such as GST or MBP), we recommend cleaving the fusion partner prior to submitting it for immunization. Large fusion partners tend to be highly antigenic, and seperate purifications are needed on the resulting anti-serum to eliminate antibodies produced against these molecules. We also recommend than any antigen with a molecular weight of less than 10kDa be conjugated to a carrier protein to make it large enough to be recognized by the host animals’ immune systems. Please contact us at abservices@prosci-inc.com if you are uncertain about any of the antigen requirements for custom polyclonal antibody production.

• Antigens in Solution: If antigens are sent in solution they should be submitted at a concentration of 1mg/mL or higher (please specify the concentration, total volume, and total amount). Precipitation is not problematic for use in immunization for custom polyclonal antibody production. We need to know the buffer components that the antigen is in (PBS, Tris, < 4M urea are preferred); SDS and citrate buffers are also acceptable. It is very important that the solution not contain detergents, as these are harmful and often toxic to the animals. Please note for services such as ELISA and immuno-affinity purification the antigen needs to be soluble, and for immuno-affinity purification the protein cannot be in Tris.

• Lyophilized Antigens: If antigens are sent lyophilized we need to know the total amount of antigen contained, if there is a preferred storage temperature (4°C, -20°C, -80°C), and if there are any specific reconstitution steps needed.

• Gel Strips as Antigens: Proteins can be submitted as a gel strip, cut from an SDS PAGE gel. Please note that we cannot perform an affinity purification or ELISA when the protein is supplied in this form. The protein needs to be reasonably concentrated (2-5 mg/mL) and then run into the gel. If it precipitates, use a lower concentration. The gel is further diluted by ProSci in 2-2.5mL of saline and 2mL of adjuvant. 100 to 200µg of protein in 500µL of gel is the maximum we can inject into rabbits. Stain the whole strip with coomassie blue (same as gel code blue stain). Then de-stain it, rinse it with water, and cut the desired strip from the gel. Do NOT “chomp” it up. After cutting the band from the gel, place it in a 15 or 50mL conical vial with an icepack – don’t freeze it. Use a fixative if desired. Please rinse twice with water to remove any remaining buffer.

  • 30% Methanol
  • 5-10% Acetic Acid
  • Fill remaining 60-65% with dH₂O

Packages:

We offer packages for client-provided antigens, single-peptide projects, peptide cocktail projects, and phospho-peptide projects.

Custom Polyclonal Antibody Production Package 1 uses a client-provided antigen, and includes an 8-week immunization protocol in a species of your choice. We can start a project with as little as 0.25mg of your antigen, or perform a complete immunization protocol with as little as 0.5mg of antigen in two chickens. We can perform additional services (ELISAs, purifications, western blotting, etc.) for any project to assist you in evaluating the antibodies produced.

Custom Polyclonal Antibody Production Package 2 includes: complimentary antigen design, synthesis of a 15-residue peptide at >70% HPLC purity, conjugation of the peptide to a carrier protein, an 8-week immunization protocol in a species of your choice, and an ELISA to verify titers on the first production bleed. A titer of >1:5,000 is guaranteed if you use a peptide sequence we approved.

Custom Polyclonal Antibody Production Package 3 includes: complimentary antigen design, synthesis of a 15-residue peptide at >70% HPLC purity, conjugation of the peptide to a carrier protein, an 8-week immunization protocol in a species of your choice, an ELISA to verify titers on the first production bleed, construction of an immuno-affinity column using project-peptide, and purification of 25mL of serum or yolk. A titer of >1:5,000 is guaranteed if you use a peptide sequence we approved.

Custom Polyclonal Antibody Production Package 4 includes: complimentary antigen design, synthesis of a 15-residue peptide at >70% HPLC purity, conjugation of the peptide to a carrier protein, an 8-week immunization protocol in a species of your choice, an ELISA to verify titers on the first production bleed, construction of an immuno-affinity column using project peptide, and purification of 200mL of serum or yolk. A titer of >1:5,000 is guaranteed if you use a peptide sequence we approved.

Peptide Cocktail Antibody Production Packages includes all of the services in Package 2 and Package 3 respectively, but include the synthesis, conjugation, and immunization of your choice of 2 or 3 peptides into one set of animals. This is an option that increases the likelihood of a peptide-based antibody working for multiple applications, as it doubles or triples the available number of epitopes for the animals’ immune systems to respond against. It also helps to reduce overall cost, as it minimizes the number of animals needed for a particular project.

Phospho-Specific Antibody Production Packages includes all of the services in Package 2, 3, and 4 respectively, but includes the synthesis, conjugation, and immunization of phospho peptides into a set of rabbits. Packages 2PN, 3PN, and 4PN also offer the synthesis of the corresponding non-phospho peptide sequence, used for isolating both the phospho and non-phospho antibodies for your target protein.

Additional services ProSci offers include:

• Peptide synthesis
• HPLC purification of peptide to >90% purity
• Conjugation of peptides and proteins to carrier proteins
• ELISA titration
• Protein A/G/L purification
• IgY purification
• Immuno-affinity purification
• Western blot analysis
• Immunohistochemistry analysis
• Project protocol extension
• Project exsanguination

Downloads:

Rabbit Custom Polyclonal Antibody Production:

Rabbit Custom Polyclonal Antibody Production Info Sheet (PDF) Rabbit Custom Polyclonal Antibody Production Order Form (PDF)

Chicken Custom Polyclonal Antibody Production:

Chicken Custom Polyclonal Antibody Production Info Sheet (PDF) Chicken Custom Polyclonal Antibody Production Order Form (PDF)

Goat Polyclonal Production:

Goat Custom Polyclonal Antibody Production Info Sheet (PDF) Goat Custom Polyclonal Antibody Production Order Form (PDF)

Background Information on Custom Polyclonal Antibody Production:

• Animal Procedures
• Rabbit Care Information
• Chicken Care Information
• Goat Care Information

If you have any questions regarding our custom polyclonal antibody production service please contact us at abservices@prosci-inc.com, or by calling toll-free 888.513.9525.