Custom Antibody FAQs
The average turnaround time from start to finish is 8-11 weeks for supplied protein antibodies, 11-14 weeks for peptide antibodies, and 12-16 weeks for monospecific antibodies. Hybridoma development requires 4-6 months to complete.
A minimum of 1mg of protein is needed to complete the Thermo Scientific, Pierce rabbit protocols (4mg is needed for the larger species such as goat), and the concentration needs to be at least 100µg/mL.
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. 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.
A total of 4 immunizations are administered to each rabbit in the 90-day protocol. Injections are given in 4 separate subcutaneous sites. A total of 4 bleeds are taken including the pre- immune bleeds. With the 70-day protocol, one less bleed is taken. The various protocols are available for review on website.
You can expect to receive approximately 90mL of sera per rabbit from the standard 90-day protocol and approximately 70mL of serum from the 70-day protocol. This does not include the exsanguinatory bleed if ordered as well.
Approximately 40-50 mL per rabbit.
Once the designated protocol has been completed, a final package will be shipped to you that includes the products and instructions for how to proceed with arranging extension or termination of your animals. You will have 10 days after the standard protocol 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.
There is no charge to terminate a project unless you order terminal bleeds.
For rabbits, we guarantee a minimum titer of 10,000 in at least one animal for any antigenic designed and made by Thermo Scientific, Pierce products. We also guarantee to follow all of the protocols as outlined and to do everything possible to induce a significant immune response and generate antibodies that work against the endogenous protein in your assay of interest. 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.
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.
Yes, complete antigenicity analysis services are offered with our advanced antigen design software called Antigen Profiler. 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).
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 2-20 mg of purified anti-immunogen antibody per 100 mL of sera.
Approximately 5mg of peptide is used for purification. If the peptide is made by Thermo Scientific, 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 us make the peptide.
All rabbits used by Thermo Scientific are assured by the Office of Laboratory Animal Welfare. Upon request additional accreditions can be provided such as AAALAC.
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 Thermo Scientific 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.
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 50,000, this means that the antibody effectively detects the antigen when bound to the solid phase at a dilution of 1:50,000.
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 the 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. The immunization schedule can be viewed when ready through OpenProject, which is the online database.
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.
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.
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.
Tools and information
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