Antigen Submission Guidelines
Pacific Immunology® provides custom antibody production services using customer-supplied antigens. To maximize the success of your project, please review the following guidelines when preparing your antigen.
Types of antigens accepted:
We accept soluble proteins, insoluble proteins, fusion tag proteins, gel bands and conjugated small molecules. We can only immunize with viral or bacterial samples that are dead and are confirmed to no longer be active.
Quantity of Antigen:
2-3 mg of the antigen is recommended. However, 1-1.5 mg of the antigen will allow for full immunization quantities if no extensions or immunochemistry services are ordered. Also, the immunogenicity of the antigen is more important than the quantity being immunized. For example, immunizations of 5-10 µg of a very immunogenic antigen can elicit high antibody titers, while immunizations with maximum quantities of a non-immunogenic antigen can elicit low antibody titers.
Antigens provided in most biological buffers in the range of pH 6-8 are acceptable. Antigens in protein solubilization reagents such as 4-8 M urea can be used, but detergents should be avoided due to interference in the production of a stable adjuvant emulsion and subsequent presentation of the antigen to the immune system.
1 mg/ml is an ideal target, but higher concentrations are welcome and most concentrations are okay if we know the approximate measurement. .25 mg/ml is the recommended lower limit.
General Ordering Information:
Please ship the antigen via FedEx or UPS overnight to:
1672 Main St. Ste. E #171
Ramona, CA 92065
Molecular Weight of Antigen:
Antigens weighing less than 6 kD are too small to elicit an immune response by themselves and need to be conjugated to a carrier protein such as KLH. Antigens weighing more than 6 kD can be immunized directly.
Fusion Tag Protein Considerations:
A protein / fusion tag complex can be used directly for immunizations. However, antibodies will be generated against the fusion tag in addition to the target protein and so may contribute to cross-reactivity in assays. Smaller tags such as His Tags are preferable to larger tags such as MBP.
Only soluble proteins can be used to coat microwells for running ELISAs and only proteins that are soluble in PBS can be used to prepare the affinity column for affinity purifications.
Gel Band Considerations:
Next: Gel Band Proteins