Custom Phosphorylation Specific pAb Production
Phosphorylation refers to the covalent modification of proteins, through the addition of a PO4 group, following completion of protein synthesis. The phosphate group is typically added at a serine, threonine, or tyrosine side chain through phosphoester bond formation.
Bon Opus Biosciences' custom antibody services allow you to entirely customize the generation of your affinity-purified phosphorylation-specific antibody. Let our team's expertise help you in the design and synthesis of your antigenic peptide. Our list of services includes consultation with an antibody expert, modified peptide antigen production, and antibody purification. When combined, these services form the basis to create a custom antibody reagent, designed with your phosphorylation site of choice, capable of performing in your desired immunoassay.
The phases of custom phosphorylated antibody generation workflow are detailed below our all-inclusive antibody production package. An antigen conjugated to KLH represents the starting point of your antibody generation process:
$1,500 Phospho and Non Phospho Antibody Package
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Includes peptide synthesis, KLH conjugation, rabbit immunization, and pAb affinity purification
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Deliverables: 2-3 mg pair of peptides, 2-5 mg phospho specific and non-phospho specific antibodies, ELISA results (1:32,000)
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10-12 week turnaround time
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1) Antigen Preparation
a) Synthesis of target peptide sequence
b) Chemical labeling of the specific phosphorylated site (Serine/Threonine/Tyrosine)
c) Conjugation of KLH to the peptide at either N- or C-terminal Cysteine
2) Antibody Production
a) Immunization of rabbits
b) Boosting of rabbits 3-4 times
c) Collection of serum
d) Testing of titers
e) Terminal bleed serum obtained
3) Antibody Purification
a) The antibody is purified through the use of a specific phospho peptide
b) Flow through and elution are differentially collected
c) Phospho specific antibodies are collected as the selected product
d) Non phospho specific antibodies are removed
The diagram below demonstrates the different antibody types produced: