Welcome to our custom antibody FAQ! If you're in need of specialized antibodies for your research or diagnostics, you've come to the right place. At Davids, we understand the critical role antibodies play in advancing scientific discovery and improving human health. Our expertise in antibody production allows us to offer tailor-made solutions to meet your specific needs.
In this FAQ, we aim to address common questions and concerns you may have about our custom antibody services. Whether you're new to the field or a seasoned researcher, we'll provide you with valuable insights into the antibody development process, project timelines, pricing and much more.
Our team of highly skilled scientists and technicians are passionate about generating high-quality antibodies that deliver reliable results. We leverage our knowledge and experience to guide you through every step of the custom antibody production journey, from antigen design and immunization to purification and validation.
By exploring this FAQ, you'll gain a deeper understanding of the benefits of custom antibodies, how they can enhance your experiments or diagnostics and the various customization options we offer. Whether you require monoclonal or polyclonal antibodies, for use in immunohistochemistry, flow cytometry, ELISA, or other applications, we've got you covered.
Q: What are custom antibodies?
A: Custom antibodies are antibodies that are specifically generated to recognize a particular antigen of interest. They are produced through a process that involves immunization, antibody screening and purification, tailored to meet the specific requirements of the researcher or customer.
Q: Why would I need custom antibodies?
A: Custom antibodies offer several advantages over commercially available antibodies. They can be designed to target unique or rare antigens, provide greater specificity and sensitivity for specific applications and offer control over antibody characteristics such as species specificity, isotype and labeling.
Q: How are custom antibodies produced?
A: Custom antibody production involves immunizing an animal, such as rabbit, chicken, mouse, rat or guineapig with the antigen of interest. The animal's immune system recognizes the antigen as foreign and starts an immune response by producing antibodies. These antibodies are then harvested, purified and validated for their specificity and functionality.
Q: What animals are commonly used for custom antibody production?
A: Commonly used animals for custom antibody production include rabbits, mice and chicken. Each animal has its advantages and considerations in terms of immunogenicity, antibody yield and production timeline. The choice of animal depends on factors such as the desired antibody type, specificity and application.
Q: What is the timeline for custom antibody production?
A: The timeline for custom antibody production can vary depending on factors such as the animal species, immunization schedule, antibody type (monoclonal or polyclonal) and the desired antibody quantity. Typically, custom antibody production can range from a few weeks to a few months.
Q: How is antibody specificity ensured during custom antibody production?
A: Antibody specificity is ensured through careful antigen design, immunogen preparation and antibody screening. Validation methods such as ELISA or Western blotting are employed to confirm that the generated antibodies specifically recognize the target antigen. With an aliquot of a test bleed customers can determine in their lab if the antibody works in their applications like WesternBlot or IHC.
Q: What are the different types of custom antibodies that can be generated?
A: Custom antibodies can be either monoclonal or polyclonal. Monoclonal antibodies are derived from a single clone of B cells and are highly specific to a single epitope. Polyclonal antibodies, on the other hand, are a mixture of antibodies produced by different B cells and recognize multiple epitopes on the antigen.
Q: Can custom antibodies be modified or labeled?
A: Yes, custom antibodies can be modified or labeled according to specific requirements. Common modifications include biotinylation, conjugation to enzymes or fluorophores. These modifications can enable various downstream applications such as immunofluorescence, immunohistochemistry, or affinity purification.
Q: How much do custom antibodies typically cost?
A: The cost of custom antibodies can vary depending on several factors, including the antibody type, production scale, modifications and labeling requirements. Use Davids Antibody Creator to receive a quote for your special needs: Antibody Creator
Q: What are monoclonal antibodies?
A: Monoclonal antibodies (mAbs) are a type of antibody produced by identical immune cells derived from a single clone of B cells. They are designed to specifically recognize a single epitope on an antigen, offering high specificity and consistency in binding. Though, compared to polyclonal antibodies, that detect multiple epitopes from one antigen, the reaction can be lower.
Q: How are monoclonal antibodies generated?
A: Monoclonal antibodies are typically generated through a hybridoma technology. This involves immunizing an animal with the target antigen, isolating B cells from the immunized animal's spleen, fusing these B cells with myeloma cells to create hybridoma cells and selecting and culturing the hybridomas that produce the desired monoclonal antibodies.
Q: What are the advantages of monoclonal antibodies?
A: Monoclonal antibodies offer several advantages, including high specificity, consistency and reproducibility. They can be produced in large quantities and have low batch-to-batch variability. Monoclonal antibodies also provide the ability to target specific epitopes, making them valuable tools for research, diagnostics and therapeutic applications.
Q: How are monoclonal antibodies validated for specificity?
A: Monoclonal antibodies are validated for specificity using various techniques, including ELISA or Western blotting. Aliquots from purified antibodies can be provided, so that you can check in your own lab if the antibodies fit for your applications like immunohistochemistry, flow cytometry, or other appropriate assays. These validation methods help confirm that the antibodies specifically recognize the target antigen and do not cross-react with unrelated molecules.
Conjugation of Antigens
Q: What is an antigen carrier?
A: An antigen carrier is a substance that is used to improve the immunogenicity of an antigen during custom antibody production. It helps stimulate a stronger immune response, leading to the production of more specific and robust antibodies.
Q: When is it necessary to use an antigen carrier?
A: An antigen carrier is typically used when the target antigen alone is not sufficiently immunogenic. This can happen when the antigen is small, lacks complexity, or is not naturally recognized by the immune system as foreign. In such cases, an antigen carrier is employed to enhance the immune response and increase the chances of generating antibodies against the desired antigen.
Q: What are some common types of antigen carriers?
A: Commonly used antigen carriers include proteins such as keyhole limpet hemocyanin (KLH), bovine serum albumin (BSA), ovalbumin and glutathione S-transferase (GST). These carriers possess immunogenic properties and can effectively enhance the immune response when coupled with the target antigen.
Q: How does an antigen carrier improve immunogenicity?
A: An antigen carrier acts as a larger, more complex structure that presents the target antigen to the immune system in a way that triggers a stronger response. It helps recruit and activate immune cells, such as B cells and T cells, which are crucial for antibody production. By conjugating the antigen to a carrier, the immune system recognizes the combined structure as a foreign invader and starts a robust immune response.
Q: Are there any considerations when selecting an antigen carrier?
A: KLH usually leads to the best results regarding the boost in the immune response. Though when the antigen is derived from a marine organism or from snails, another carrier might be more suitable.
Q: Can an antigen carrier affect the specificity of the generated antibodies?
A: In some cases, the use of an antigen carrier can induce an immune response against both the carrier and the target antigen. This may result in the production of antibodies that recognize both the carrier and the antigen. For this reason Davids recommends to perform an affinity purification with an unconjugated antigen when it comes to peptide antibodies for example. Another way to get rid of unwanted antibodies is a depletion.
Q: Do I always need an antigen carrier for custom antibody production?
A: Not necessarily. Some antigens may already possess sufficient immunogenicity and can elicit a robust immune response without the need for an additional carrier. However, when working with antigens that are known to be poorly immunogenic or have a low molecular weight, incorporating an antigen carrier is often recommended to improve the antibody generation process.
Purification of Antibodies
Q: Why is antibody purification necessary?
A: Antibody purification is essential to isolate and obtain highly pure antibodies from complex biological samples or production systems. Purification removes unwanted contaminants, ensuring the specificity and functionality of the custom-made antibodies.
Q: How are custom antibodies purified?
A: Custom antibodies can be purified using various techniques, including protein A/G/L affinity chromatography, ion exchange chromatography, size exclusion chromatography and affinity purification using antigen-specific columns. The choice of purification method depends on factors such as antibody type, scale and downstream application requirements.
Q: What is protein A/G/L affinity chromatography?
A: Protein A/G/L affinity chromatography is a commonly used method for antibody purification. These protein ligands bind to the Fc region of antibodies, allowing for selective purification. Protein A is specific to IgG subclasses, protein G binds to a broader range of species and IgG subclasses and protein L is specific to certain IgG subclasses. The result is a whole IgG fraction with specific and unspecific antibodies.
Q: What is ion exchange chromatography?
A: Ion exchange chromatography separates antibodies based on their net charge. It utilizes a stationary phase with charged groups, either positively (cation exchange) or negatively (anion exchange) charged, to attract and bind antibodies with opposite charges, enabling their separation from other molecules.
Q: What is size exclusion chromatography?
A: Size exclusion chromatography separates antibodies based on their size or molecular weight. It employs a porous matrix that allows smaller molecules to enter the beads and elute later, while larger antibodies elute earlier due to their limited access to the pores. With this method dimers or IgM can be seperated from IgG.
Q: Can affinity purification using antigen-specific columns be used for custom antibody purification?
A: Yes, affinity purification using antigen-specific columns can be employed to purify custom antibodies. The custom antigen is immobilized on a solid support, allowing the specific antibodies to bind, while contaminants are washed away. Elution of the purified antibodies is typically achieved by altering the pH or using competitive agents. The results are specific antibodies against the antigen.
Q: Are there any considerations for downstream applications during antibody purification?
A: Yes, downstream applications can influence the purification strategy. If the custom antibodies will be used for functional studies or in vivo applications, additional purification steps or removal of endotoxins may be required to ensure optimal performance and safety.
Q: Can custom antibodies be labeled or modified during or after purification?
A: Yes, custom antibodies can be labeled or modified during or after purification, depending on the specific requirements. Common modifications include conjugation to fluorescent dyes, enzymes, biotin, or other molecules, enabling various detection or applications.
Q: What is the typical yield of purified custom antibodies?
A: The yield of purified custom antibodies can vary depending on several factors, including the starting material, purification method and antibody characteristics. If you have further questions regarding the estimated yield, please let us know.
Storage of antibodies
Q: How do you ship the antibodies?
A: Davids ships their antibodies with cool packs. Please note that the antibodies are stable at room temperature for several days. Even when the parcel is delayed the antibody acitivity is not affected.
Q: How do I store my antibodies after arrival?
A: Davids highly recommends to store the antibodies at 2 - 8°C.
Q: Can I freeze the antibodies?
A: Davids does not recommend to freeze antibodies. Each freezing/thawing cycle can reduce the activity of the antibodies. When you want to store them at -20°C or even -80°C, we recommend that you try with a small aliquot if the acitivity of the antibodies remain after one freezing/thawing cycle.
Q: Are there any preservatives in the antibody solution?
A: Davids adds Sodium-Azide to the antibodies as preservative. When you do not want a preservative please let us know at the start of the project and we can discuss an alternative storage buffer.
Q: How can I order custom antibodies?
A: The order process is rather simple. We recommend that you get your own quote ( Antibody Creator) and send this quote together with your antigen to Davids.
Q: How can I order peptide antibodies?
A: When you send us your amino acid sequence from your protein of interest by email, Davids will suggest you peptides that can represent your protein. When you want to continue with the order, you can just confirm the peptides by email and we start with the synthesis.
Q: Do I need a PO number?
A: Davids does not need a PO number. Though many companies and universities need a PO number on the delivery note and the invoice to connect the order with the delivery. In this case please provide us your PO and we can include it in the order.
Confidentiality and Restrictions
Q: Can I use the antibodies to make a patent or a publication?
A: After payment of the antibodies the antibodies are yours and you can use them for your scientific work and make your publication.
Q: Are my data handled confidential?
A: We know that all scientific data is important. For this reason we handle all data from our customers confidential. This includes sequences, proteins names and information about our customers.
Q: Where can I find references about Davids antibodies?
A: As mentioned earlier, Davids handles all customers confidential. For this reason, we cannot give references. But you may find publications that used Davids antibodies in the Internet. Try to search for: "davids biotechnologie" in famous databases.