FAQs

GENERAL

Does Pacific Immunology® guarantee antibody titers?

Yes, Pacific Immunology® guarantees titers of at least 1:100,000 against peptide sequences that we recommend / approve, synthesize and conjugate. Titers are confirmed via ELISA at the 1st production bleed.

Does Pacific Immunology® offer any catalog antisera?

No, to avoid conflicts of interest, Pacific Immunology® focuses exclusively on the production of custom antisera for its clients.

Do I retain rights to the antisera that I generate?

Yes, clients maintain full IP ownership of custom antibodies.

Does Pacific Immunology® maintain the confidentiality of my project?

Yes, Pacific Immunology® has for decades emphasized the importance of safeguarding clients’ confidential information.

Does Pacific Immunology® generate Monoclonal antibodies?

Yes, Pacific Immunology specializes in the development of Rabbit Monoclonal antibodies with proprietary screening technologies to quickly and cost effectively deliver sequence-defined, recombinant monoclonal antibodies.

What is the advantage of Pacific Immunology®'s AdjuLite™ Freund's Adjuvants?

Pacific Immunology®'s ultra pure Freund's Adjuvants are comprised of premium components that enhance the immune response and significantly reduce immunization stresses for the animal. Please visit our AdjuLite™ Freund's Adjuvant page for additional details.

Does Pacific Immunology® offer volume discounts?

Yes, we offers discount for starting multiple projects at the same time.

What is Sodium Azide and should I add it to the antiserum?

Sodium Azide is a preservative that prevents bacteria from growing in the serum (bacteria can produce proteases that denature any proteins in the serum, including antibodies).

How will the serum be shipped?

Serum is typically shipped via FedEx Overnight immediately after each bleed is taken.

What yields of purified antibody can be expected?

A typical affinity purification of 25ml of serum will isolate approximately 2-3mg of peptide-specific antibody.

Will antibody titers increase with additional immunizations?

Antibody titers typically stabilize after the 1st production bleed with additional immunizations generally helping maintain (rather than increasing) antibody titers.

Are accelerated protocols available?

No, accelerated protocols are a marketing gimmick that delivers inferior, lower affinity antibodies.

What kind of rabbits / chickens does Pacific Immunology® use?

New Zealand White Rabbits and Rhode Island Red Chickens

Will I receive any leftover peptide at the end of the project?

Yes, you will typically receive at least 30 mg of standalone peptide at the completion of the project.

ANTIGEN PREPARATION

How much protein do I need to send?

We recommend sending at least 1-2 mg of the antigen for a rabbit or chicken project and 5 mg for a goat project. We recommend a concentration of at least .5 mg/ml if possible.

How should I send my protein to Pacific Immunology®?

Please review our Antigen Preparation guidelines

Does the protein need to be soluble?

No, insoluble proteins can be used for immunizations.

Can I generate antisera against gel strips containing my protein?

Yes, immunizing with gel bands can work well for generating polyclonal antisera against a purified protein. Please review our Antigen Preparation guidelines

Should I generate antibodies against a full-length protein or a peptide?

Targeting full-length proteins allows for the development of antibodies against multiple epitopes throughout the protein, thereby increasing the probability of recognizing the native protein in the target assay. However, specificity is typically compromised for polyclonal projects. Expression of the protein may also be difficult or prohibitively expensive. Targeting peptide sequence can be a cost effective method to quickly isolate antibodies with high specificity to the target protein; but, binding capability with the native protein may be compromised in some assays.

Does Pacific Immunology® offer antigen design assistance?

Yes, our peptide chemists will analyze your protein sequence using an advanced suite of algorithms to recommend the most immunogenic sequences with a high probability of exposure.

Why is conjugation of the peptide to a carrier protein necessary?

The molecular weight of most standalone peptides is too low to generate an immune response in the animal.

What role does the adjuvant play and what is the difference between CFA and IFA?

An adjuvant is a substance that, when combined with the antigen, serves to enhance the immune response against the antigen. Freund's adjuvants are the preferred adjuvant of choice for use in antibody production, and Pacific Immunology® uses its own premium line of AdjuLite™ adjuvants for all immunizations. AdjuLite™ Complete Freund's Adjuvant is used only for the first immunization and contains mycobacteria as an additional stimulate for the host's immune system. AdjuLite™ Incomplete Freund's Adjuvant is used for all subsequent immunizations. Please visit AdjuLite™ Freund's Adjuvant for additional details.

ANTIBODY USAGE

How should I store the serum / antisera?

Please review our Antibody Storage guidelines

Why am I seeing multiple or unexpected bands on a Western with my affinity purified antibody?

Please review our Western Blot troubleshooting guidelines

Why am I not seeing any bands on my western when assaying with the purified antibody?

Please review our Western Blot troubleshooting guidelines

How do I interpret the ELISA results?

The ELISA measures the peptide-specific immune response by comparing dilutions of the pre-immune serum and the first production bleed serum. A titer of 1:50,000 to 1:100,000 generally indicates a strong, specific immune response.

PURIFICATION

What is the difference between an IgG purification and an affinity purification?

An IgG purification isolates all IgG in the serum, regardless of the specificity. This is most often used for projects targeting full-length protein where affinity purification may not be possible. Affinity purification, by contrast, is advantageous for isolating only those antibodies that recognize a small subset of epitopes (typically peptide targets), helping approach the specificity of monoclonal antibodies at a reduced cost.

Is the affinity column included so that I can run additional purifications?

Yes, the affinity column is included for purifying additional lots of serum.