Custom Polyclonal Antibody Production Services
Biomolecular Integrations offers a one-stop solution for all your polyclonal antibody production, purification and characterization needs. We are specialized in generating high-affinity, high titer and specific polyclonal antibodies to your target antigens using our standardized as well as customized antibody production protocols that suit your requirements. We will discuss your project personally to know your requirements and then advise you of an effective strategy to obtain the kind of antibody that you need. We can generate antibodies in any species of your choice including rabbit, goat, sheep, horse and chicken. Polyclonal antibodies can be delivered as serum or affinity purified as per your requirement. We also generate phosphospecific antibodies.
Free Peptide/Antigen Design Service
Biomolecular Integrations offers free peptide antigen design service to help you design the optimized peptide for your antibody experiments. We perform protein databse BLAST, hydropathy and antigeneicity analyses for the selection of most suitable antigenic peptides. This will minimize solubility problems and maximize your chances of obtaining the high titer antibody. Our PhD level scientist will provide you customized consultation, detailed reports and sequences of the peptides in easy to understand format. ...
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Peptide Synthesis
All peptides used for antibody production are synthesized by our expert service providers. The peptide purity may vary depending on the size and complexity of the peptides. We offer competitive and fixed prices with no set up fee on peptide synthesis. Generally our peptides are greater than 80% pure and at a scale higher than 20 mg for antibody production purposes. The quality is determined by HPLC purification, gradient elution, massspectrometry analysis and UV scans. Post synthesis peptide modifications such as phosphorylation, cyclization, terminal amidation, farnesylation, sulfation etc. are extra. Peptides with higher purity (<98%) and larger scale can be synthesized at an extra cost...
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Customer Supplied Antigens
We can also assist you if you would like us to produce antibodies against your own supplied proteins (i.e. gel purified protein, fusion protein, recombinant protein, or others). For the Standard rabbit protocol, please supply approximately 5 mg protein at a concentration of at least 0.5mg/ml. Customers can supply proteins as lyophilized powder or in solution without organic solvents. ...
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Peptide Conjugation
Peptide conjugation service is available as per customer requirement. Customers may choose various carriers such as keyhole limpet hemacyanin, (KLH), bovine serum albumin (BSA) and ovalbumin (OVA) or we may use our own conjugation methodology most suitable for better antigen presentation. ...
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Antibody Purification
The highest level of purification is achieved with our peptide-affinity chromatography, which is unmatched in producing high affinity and functional antibodies. We offer selection of mono-specific polyclonal antibodies by successive batch elution over multiple affinity matrices and phospho-specific antibodies by selectively isolating on phospho-peptide affinity columns. We also offer Protein A, G or L purification and cross-adsorption services. All purified antibodies are analyzed with ELISA and absorbance. ...
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Antibody Screening/ testing
Biomolecular Integrations, Inc. offers ELISA screening of antibody titers by coating free peptides or peptides coupled to a carrier protein other than used for immunization using our optimized ELISA protocols. This way, antibodies to carrier proteins are not detected. We perform ELISA on individual bleeds along with pre-immune serum at 5 dilutions (1:10 , 1:100, 1:1K, 1:10 K and 1:100K). The ELISA results are tabulated in an easy to understand format. Phospho-peptide antibodies are characterized by ELISA using phospho and dephospho peptides also at 5 dilutions. The Phospho-specific antibodies are later isolated by sequential affinity purification on dephospho and phospho peptide affinity matrices. The detailed methodology for ELISA is provided with each antibody progress report for easy reproduction of results in investigators laboratory if needed. All ELISA reagents are also available from Biomolecular Integrations, Inc. We also characterize antibodies by Western Blot analysis. ...
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Other Services
Besides our free consultation and support services for antigenicity determination, peptide design, antibody purification and characterization, following optional services are also available at competitive prices.
· Synthesis of peptide antigens
· Antigen conjugation with career proteins
· Antigen emulsification in Freund's adjuvants or the adjuvant of your choice
· Sera testing and titer determination
· Affinity purification with specific peptide or protein A/G affinity columns
· Customized antibody labeling with preferred tag
· Antibody characterization by ELISA, Western blotting, immunohistochemistry, immunocytochemistry)
· Development of custom screening methods
· Antibody fragment preparation: Fab or F(ab)'2
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Request a Quote or Place an Order
Please submit your inquires via email technical@biomolecularint.com or call our PhD level antibody experts at 501 247-3248 with your specific questions. You may also submit your inquiry online. By learning more about your project and specific requirements, we may be able to help you design better antigens and plan an appropriate strategy for your specific project. We will also provide you with an accurate price quote & timeline for your project with no obligation to buy. You may also call us directly at 501-247-3248 to place an order or fax your order at 501-661-0202, or email us at customer@biomolecularint.com.com with your purchase order as attachment. We accept credit cards or purchase orders from established institutions....
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Guaranteed Service
Biomolecular Integrations offers money back guarantee on polyclonal antibody production. This means that you will not be charged if we are not able to produce antibody to your antigen designed by us. We guarantee a 1:10,000 ELISA titer in the last production bleed or we will re-do the protocol for free. We have a good success rate of developing antibodies to a verity of antigen types. However, due to inherent risks associated with antibody development, there is no absolute guarantee to get the desired antibody. A number of factors may affect antibody production but we try our best to ensure that success is achieved. Your investment is protected at Biomolecular Integrations in an event of unforeseeable problem which means that only start-up fee and costs associated with antigen preparation will be billed. The remaining portion of the project will not be charged. The entire amount of the project will only be charged upon successful completion of the project. Obviously there is always some loss of time in such unlikely scenario. Biomolecular Integrations is not responsible for any loss of time.
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Standard Rabbit Protocol for Protein and Peptide Antigens
Day 0 Pre-immune bleed collected and stored at -20 0C until the first test bleed. Immunization with 100-200ug protein in Complete Freund's Adjuvant (CFA).
Day 14 First booster injection with 50-100ug protein in Incomplete Freund's Adjuvant (IFA).
Day 28 Second booster injection with 50-100ug protein in IFA.
Day 42 Test bleed. The pre-immune bleed and this first test bleed will be shipped to the customer within the next two days for screening of the antibody titer until antibody titer determination service is requested. We offer antibody titer by ELISA.
Day 56 Test bleed. Third booster injection with 25-50ug protein in IFA
Day 70 Production bleed. We will continue to bleed every 14 days unless customer specifies after determining antibody titer and the quality of antibodies, and let us know if any changes are necessary in the protocol.
Day 84 See for continuation of your protocol below.
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Standard Rabbit Protocol for Fusion Protein Antigens
Day 0
Pre-immune bleed collected and stored at -80 0Cuntil the Day 63 bleed. Immunize with 100-200ug protein in Complete Freund's Adjuvant (CFA). Acrylamide protein extracts can only be injected subcutaneously.
Day 28
First booster immunization with 50-100ug protein in Incomplete Freund's Adjuvant (IFA)
Day 42
Second booster injection with 50-100ug protein in IFA.
Day 56
Test bleed. Thirst booster injection with 25ug protein in IFA. Inject 25ug protein intravenously (antigen must be soluble in PBS or water) using no adjuvant. If this is not available, we will skip the intravenous injection.
Day 63
Production bleed. We will ship pre-immune bleed, the Day 56 test bleed and this production bleed. The investigator tests the sera, determines titer and the quality of antibodies, and informs us if for any changes in the protocol. We also offer antibody titration service by ELISA.
Day 70
Fourth booster injection with 25-50ug protin in IFA.
Day 84
Test bleed. We will do these bleeds every 14 days unless the investigator specifies.
After Day 84
See Continuation of your Protocol below.
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Continuation of Protocols
All protocols are continued by giving booster injections every four weeks and test bleeds two weeks after each booster injection. The serum is shipped within two days of bleeds. You have option to stop or continue your protocol depending upon your needs and the success of the project. When an antiserum gives sufficient antibody titer, we start production bleeds every two weeks. We can also do production bleeds every week. Please contact us if you want to change the bleeding and/or boosting schedule. You may also request that we do not do any booster injections after the initial immunization outlined above since some antigens and animals may respond negatively to boosts during the period of peak antibody activity. Boosts may be required only when titer drops. It is up to the investigator to decide and inform us whether to discontinue boosts. The volume of serum collected for large or production bleeds depends on the weight and the general health of the Rabbit. We generally obtain 20-40ml blood yielding 10 to 20ml of serum.
In our experience, the peak antibody titer occurs at an average of 120 days, however, the onset of peak may occur as early as six weeks and may last as long as one year. If the results are not as you expect, please consult us for recommendations on how to modify your protocol. When you have collected enough antisera, you will need to contact us to terminate the project. You may request that we perform exsanguinations (terminal bleed) or if no additional sera are required, the animal(s) will be humanely euthanized (discard). When you instruct us to terminate your animal(s), you must tell us if you want exsanguination or discard, species, and the animal(s) number. Written confirmation by email or fax may be requested for terminal procedures. ...
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