1. How long does it take to get my antibody?
   The average turnaround time from start to finish is 11 weeks for supplied protein antibodies, 14 weeks for peptide antibodies, and 16 weeks for monospecific antibodies.

2. If I want to raise antibodies to a recombinant protein that I have, how much protein do I need to send and at what concentration does the protein need to be?
   A minimum of 1mg of protein is needed to complete the Open Biosystems standard protocols (2mg is needed for the Ivan-Baines protocol), and the concentration needs to be at least 100µg/mL.

3. Can I send gel-slices containing my protein for antibody production?
   Yes, antibodies can be produced to gel slices. Cut out the relevant protein after staining and send it in a 50mL conical tube with a small amount of buffer. The buffer will keep the gel from drying out. Open Biosystems will extract the protein for immunization, but it is important that you quantitate the total protein amount as accurately as possible as there is no way of determining the protein concentration once the sample is received. You may also want to consider sending some of the non-gel form of the protein (200µg) as denatured proteins sometimes do not stick well to ELISA microtiter plates.

4.  How many boosts and bleeds are performed?
   A total of 4 immunizations are administered to each rabbit. Injections are given in 4 separate subcutaneous sites. A total of 4 bleeds are taken including the pre- immune bleeds.

5. How much serum can I expect?
   You can expect to receive approximately 90mL of sera per rabbit from the standard protocol. This does not include the exsanguinatory bleed.

6. How much serum do I get from a terminal bleed?
   Approximately 45mL per rabbit.

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7. When do I have to decide about the termination of the production animals?
   Once the designated protocol has been completed, a final package will be shipped to you that includes the products, ELISA results, and instructions for how to proceed with arranging extension or termination of your animals. You will have approximately 2 weeks to make a decision on whether you want to extend the animals with a selected extension protocol, have terminal bleeds taken on the animals, or simply terminate the project with no further action.

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8. Is there a charge for terminating a project at the end of the 10 week protocol?
   There is no charge to terminate a project unless you order terminal bleeds, which are $50 per animal (additional charges apply for some species).

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9. Do you guarantee a minimum titer?
   We do not guarantee a minimum titer. Since the immune response of the animal is dependent on the animal and the antigenicity of the peptide, we cannot guarantee final titer. We do guarantee to follow all of the protocols as outlined and to do everything possible to induce a significant immune response. ELISA titers are an arbitrary measure of antibody reactivity, which may or may not translate into the ultimate success of the antibody. Although titers do very often correspond with the performance of the antibody, we recommend using titers only as a measure of how to proceed with appropriate boosting or sera collection.

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10. Do I have to order purification in advance?
    No. Purification may be ordered at anytime. If you do place your order up front, you will only be sent aliquots of the post-inject sera instead of the bulk bleeds, which we will hold at our facility. This helps you to avoid having to ship them back to us if you decide that you do want to purify. You are not committed to purifying even if you order it up front.

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11. Can you help me to design my peptides? Do you charge for this service?
    Yes, complete antigenicity analysis services are offered. There is no additional charge for this service, and it is recommended that consultation is requested unless you are an extremely experienced producer of antibodies or you have your own internal bioinformatics program(s).

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12. How much purified antibody can I expect to receive?
    Yields from affinity purification can vary quite drastically because of antibody concentration in serum and the affinity of the anti-immunogen antibodies. As a general rule, you can expect to receive between 2-25mg of purified anti-immunogen antibody per 100mL of sera.

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13. How much peptide do you use for purification?
    7-8mg of peptide is used for purification. If the peptide is made by Open Biosystems, it is provided at no cost. If the peptide is supplied by you, you will need to send enough for the purification. Alternatively, you can have the peptide made at mid scale for $18 per residue.

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14. Is Open Biosystems AAALAC accredited?
    Open Biosystems' animal facilities are AAALACC accredited.

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15. Is prescreening rabbits necessary, and how long do I have to screen them if I decide that I do need to?
    Most customers do not prescreen rabbits prior to immunization, but this can be extremely important if you are studying certain organisms such as Drosophila. All rabbits used for antibody production by Open Biosystems are certified specific pathogen-free (SPF), which provides a cleaner basis for antibody production. If you do decide to prescreen, you are provided with two weeks from the time you receive the pre-bleeds to respond with the selected animals for immunization.

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16. How do I interpret my ELISA results?
    Crude ELISA titers are reported as the reciprocal of the serum dilution. Titers are reported this way for more convenient reporting and formatting. If you have a titer of 50000, this means that the antibody effectively detects the antigen when bound to the solid phase at a dilution of 1:50000.

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17. After I placed my order, I received a confirmation of order, but I did not receive an immunization schedule. Can you send me one?
    Detailed immunization schedules are not sent to customers until the peptide synthesis and conjugation have been completed and the rabbits have been immunized. It is done this way, because the exact immunization dates are not known beforehand due to different variables in peptide synthesis time. One of our veterinarians must also sign off on the continuation of the protocol after observing the animals following primary immunization assuring there are no adverse reactions to the antigen.

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18. I ordered three antibodies, but I only received an immunization schedule for two of them? What happened to the other antibody?
    Projects are started as soon as possible after the initial order. If one or more peptides from a multiple project order is completed and conjugated before the others, these peptides will not be held to wait on the others. The goal is to deliver each and every antibody as quickly as possible.

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19. Is MAP better than KLH?
    MAP is not better than KLH. It is simply an alternative carrier strategy that does not require the peptide to be post-synthetically conjugated. MAP peptides are recommended only for shorter peptides that are located at the n-terminus or the internal portion of the protein, while KLH-peptides are recommended for any peptide from any location within a protein. It is also a good idea to use KLH instead of MAP when there is a cysteine located in the peptide sequence.

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20. I have a cysteine located in the middle of my peptide sequence. Should I be concerned about disulfide bonding? Should I change it to another residue such as serine?
    No, you should not be concerned about disulfide bonding, and you should not change the residue. It can actually be advantageous to have an internal cysteine present.