production - an overview

Antibodies that work - whatever the application: At phosphosolutions we produce custom-made phospho-specific antibodies to the phosphorylation sites of interest to research scientists. We are dedicated to making antibodies that work, whatever the application. For a detailed quote for our custom phospho-specific antibody production service, contact us at mdb@phosphosolutions.com. A brief description of our procedures and protocols can be found below.

custom antibodies

Custom Development of Phospho-specific Antibodies: The first step in the development of custom-made phospho-specific antibodies is the selection and design of the immunizing peptide. This is a crucial step because the specificity and utility of the custom phospho-specific antibodies depends critically on the design of the immunizing peptide.

experience

Drs. Andrew Czernik and Dr. Michael Browning, who together have more than three decades of experience in designing and producing phospho-specific antibodies, are responsible for the design of the immunizing peptides at PhosphoSolutions. As an example of this process, the phosphopeptide for serine-40 in tyrosine hydroxylase is shown in Figure 1. 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 of Figure 1.

figure 1

rationale

The rationale for the design of this peptide is as follows:

Truncated N-Terminal sequence: First, the peptide has a truncated N-terminal sequence. This was done to eliminate cross reactivity with homologous proteins that were identified in a Blast search.

Use of Short Sequence: Second, another feature of the peptide design was the use of a very short sequence which forces the phosphoseryl residue into the epitope recognized by the antibody.

Amidation of C -Terminus: Third, given the very favorable amphipathic character of the peptide, the C-terminus was amidated to better emulate the native tyrosine hydroxylase protein. Such modifications can improve immunoreactivity in western blots and especially in immunohistochemical applications.

Addition of N - Terminal Cysteinyl Residue: Lastly, an N - terminal cysteinyl residue was added for conjugation to the carrier protein and coupling to the affinity column.

>>> NEXT: Antibody Purification >>>

production | purification | characterization

PhosphoSolutions

Why Go Anywhere Else? As this summary indicates, we devote a great deal of care to the production of phospho-specific antibodies for each individual research scientist's protein of interest. As you visit our site, you can learn more about our custom phospho-specific antibody production service, find out more about our scientists and how to contact us.