Custom Antibodies - Your Way! Whether you require a single protein antibody, one or more phosphospecific antibodies, or reagents to assess changes in a variety of epigenetic modifications, our scientific staff can help design robust antibodies with unsurpassed specificity. Utilizing a variety of novel and proprietary approaches no other company can match our science, our results and our customer service. • Complete protocols – from peptide to purified antibody in many different species • We make the peptides so pricing is reasonable • AAALAC/USDA/NIH/OLAW accredited farm • Final antibodies can be supplied in many forms: aliquotted, lyophilized, biotinylated, dye-labeled Modification State-Specific Antibodies When it comes to making antibodies that target post-translational modifications, our scientists tailor a specific protocol that meets your needs. •Phosphospecific (Ser/Thr/Tyr) •Epigenetic (Lys/Arg, methyl, acetyl) •Nitrotyrosine •Acylation •Glycosylation Many Options to Fit Your Needs! Before initiating an antibody protocol, you will speak with our scientists so that the antibody you receive is the antibody you need. Our scientific staff has been designing and manufacturing antibodies for nearly 30 years so whether you need an antibody for westerns, immunohistochemistry, flow cytometry, IP or other applications, we have protocols and the scientific understanding to give you the best chance to achieve your scientific goals. Complete Protocols to Meet Your Needs Research dollars are tighter than ever so we have designed flexible antibody protocols that can be tailored to meet your specific needs. Each protocol begins with the most important part: consultation with our scientists who have decades of experience in the design, execution and troubleshooting of custom antibody protocols. We achieve a success rate of over 90% across the total spectrum of antibodies we manufacture - with western blots as the minimally acceptable criteria not by guesswork and hope, but through the use of methods for each step of the process that have been carefully thought out, developed with decades of hard work and collaborative efforts with customers all over the world. Each of our protocols includes: •Peptide design and manufacture •Conjugation to carrier proteins •Immunization of animals housed at an AAALAC/USDA/OLAW accredited facility •Species for antibody production includes rabbits, goat, sheep, rodents, chickens and others •Immunization protocols that are optimized to generate sufficient antibody to meet the goals of your research •Dot blots of serum and final antibody; western blotting available as an option •Affinity purification, immunodepletion (when required), biotinylation/dye labeling, crosslinking, and antibody fragmentation available •Expert troubleshooting of difficult protocols including the use of proprietary methods to generate specific antibodies to unusual targets One of the most critical aspects of antibody production is affinity purification of the specific antibody desired. All of the hard work that goes into design of appropriate peptides or recombinant proteins can be wasted if the final product is not specific for the intended target. When producing antibodies to modified amino acids (post-translational modifications, PTMs), purification and immunodepletion of antibody not specific for the modified state of the antibody is even more critical. When working with the scientists at 21st Century Biochemicals, you are working with the people who have actually “written the book” as our Founder, Dr. Jordan Fishman and our VP of Operations, Dr. Eric Berg, wrote two chapters in the newest Cold Spring Harbor Laboratories Press book: Antibodies, A Laboratory Manual (2014), on Antibody Purification and Storage as well as Antibody Labeling. The chapter on purification gives numerous examples of how to properly purify antibodies that detect proteins in general, or antibodies that target single to multiple PTMs in the same immunogen. Antibody Purification and Storage. Jordan B. Fishman and Eric A. Berg. In: Antibodies: A Laboratory Manual, Second Edition (Edward A. Greenfield, ed). 2014 Cold Spring Harbor Laboratory Press, Chapter 10, pp. 371-404. Labeling Antibodies. Eric A. Berg and Jordan B. Fishman. In: Antibodies: A Laboratory Manual, Second Edition (Edward A. Greenfield, ed). 2014 Cold Spring Harbor Laboratory Press, Chapter 12, pp. 429-468. Frequently Asked Questions What species should I choose for generating my antibody? Based upon economic factors and the ability to generate a robust immune response in a reasonable amount of time (65-110 days of animal time), rabbits are the go to species for most custom antibody protocols. Rabbits also have the advantage of providing reasonable sized production bleeds (16-25ml/bleed) as well as an antibody- generating timeframe that provides for many months of continued boosting and bleeding. With a small cohort of good producing rabbits, hundreds of mg to even grams of affinity purified antibody can be obtained. For applications requiring multi-gram quantities of affinity purified antibody, larger animals can be used such as goat or sheep. The use of alternate species is also important in research studies where multiplexing is required – the detection of multiple targets at the same time utilizing two or more dyes emitting fluorescent signals at multiple wavelengths. What Types of Antigens Can Be Used to Manufacture a Polyclonal Antibody? Antibodies can be made from expressed or native protein, proteins separated by gel electrophoresis and the appropriate band excised, chromatographically purified tryptic peptides, heat or formaldehyde- inactivated viruses, and synthetic peptides. The use of synthetic peptides, owing in part to their minimal cost and purity, have become the most common type of antigen used for custom antibody production. Given the critical nature of the immunogen, we manufacture 100% of the peptides we sell or utilize in our antibody protocols, and we have the most rigorous QC in the industry - including tandem MS to confirm the actual sequence of every purified peptide - to insure 100% peptide fidelity. In addition, during the synthetic process, PTMs can be added at specific site(s) along the peptide chain, allowing for the production of antibodies selective not only for the peptide's primary sequence, but for one or more PTMs as well. Antibodies to numerous PTMs have been manufactured successfully by our staff, including antibodies to: •One or more phosphoserine, phosphothreonine, and/or phosphotyrosine sites •Phosphohistidine •Epigenetically modified lysine (me1, me2, me3, acetyl) •Epigenetically modified arginine (monomethyl - MMA, symmetric dimethyl - SDMA, asymmetric dimethyl - ADMA) •Hypusinated lysine •Nitrotyrosine •Oxidized methionine The best way to have all of your questions answered is to speak directly with one of our scientists.