Custom Antibody Services
Custom Polyclonal Antibodies that Work!
We will be with you through the entire process of making and testing your custom polyclonal antibody. Dr. Michael Browning, who has more than three decades of experience in designing and producing phospho-specific antibodies, is responsible for the design of the immunizing peptides at PhosphoSolutions. In addition, we are experts in purification and characterization of polyclonal phospho antibodies. We do so much more than collect and send you serum from your rabbits. We screen the serum ourselves, looking for the highest titer bleeds. We then purify the antibody over affinity columns made with the immunizing peptide, and characterize the antibody in ELISA to check for phospho specificity. After you receive your antibody for testing, we continue to work with you to optimize your signal and success.
For a detailed quote of our custom polyclonal antibody production service, contact us at info@phosphosolutions.com.
Design
The crucial first step in custom antibody development is peptide design. The specificity and utility of the antibody depends critically on the immunizing peptide. As an example of the peptide design process, consider figure below, illustrating the phospho-peptide for serine-40 in tyrosine hydroxylase. The sequence N-terminal and C-terminal to the phospho-serine is shown at the middle of the figure, while the peptide design is shown at the bottom.
Purification
After immunization with the phospho peptide, three types of antibodies can be produced in the rabbit’s immune response. As seen in Figure 1 below, the first is the phospho-specific antibody that is desired. However, antibodies that are specific for the dephospho form of the peptide can also be generated if phosphatases in the rabbit dephosphorylate the peptide conjugate that had been injected.
Characterization
The composite Western blot below in Figure 1 shows the characteristic doublet of our synapsin pan antibody as well as three of our phospho synapsin antibodies, and demonstrates two very import aspects of antibody specificity. First, there are no cross-reacting bands labeled by the antibody in the rat brain homogenate.
