Phosphorylation can play an important role in the regulation of protein activity and their stability. Phosphospecific antibodies can recognize the status of the phosphorylation site. Our phosphospecific antibody service includes all steps for the creation of your antibodies:
After synthesis and conjugation of the phosphospecific peptide (1) the animals are immunized (3) with the conjugated phospho-peptide (2) to produce antibodies. After antibody production the serum is affinity purified with the phospho-peptide to get specific antibodies (4). In this stage of the process there are still some antibodies recognizing the non-phosphopeptide. To get rid of these unwanted antibodies a depletion is necessary. The affinity purifies serum is depleted with a non-phosphopeptide depletion matrix. The result are phosphospecific antibodies (5). Prices and immunization options for phosphospecific antibodies from rabbits can be found here: Rabbit Antibodies.
Our scientists help you to to find peptides that represents your phosphospecific antigen. Our analysis is based on the solubility, the antigenicity, the epitopes and our experience. Send us your protein sequence as one-amino-letter-code and the labeled phosphorylated amino acid. We suggest you peptides that are suitable for your project. An order is only placed when you confirm the peptides.
After your order we begin with the synthesis of the phosphorylated and the non-phosphorylated peptide. The phosphorylated peptide is conjugated to a carrier to enhance the antigenicity. The non-phospho-peptide is used for the purifications.
You will receive an aliquot of both peptides for your own research.
The conjugated phosphopeptide is used for the immunizations. Our complete service A061 includes the immunization of 2 rabbits. We recommend to immunize 2 rabbits, as we remove a lot of unspecific antibodies in the affinity purification and depletion steps.
After 3 immunizations we take a test bleed to determine the titer. You can receive an aliquot of this test bleed to determine the titer in your own lab. After 2 additional immunizations within our 63-day protocol, we take the final bleed. Additional immunizations are usually not needed. Though you can order them if wanted.
The affinity purification with the phosphopeptide collects all peptide-specific antibodies. In this fraction are antibodies that recognize the phosphopeptide, but also some antibodies that recognize the non-phosphopeptide. To get rid of the last ones we perform a depletion with the non-phosphopeptide. The results are phosphospecific antibodies.
The same process of phosphospecific antibodies can be performed with other amino acid modifications like Acetylation, Glycosylation and more.
Davids keeps the order process very simple. Just send your peptide sequence to email@example.com. All data is handled confidentially. If you want to provide the peptides you can send them to the following address:
Davids Biotechnologie GmbH
We start with the immunizations as soon as the synthesis finished. Please provide us with the following information: