Post-translational modifications (PTMs) play important roles in regulating the biological activity of a protein. Phosphorylation is one of the most common PTMs carried out by the protein kinases. A properly designed P-site™-specific antibody allows one to monitor the kinase activation event without the use of radioisotope 32p.
Our team of scientists has derived a wealth of experience over the years by making hundreds of P-site™-specific antibodies (including methylations (me1, me2, and me3), acetylations, hydroxylations, neo-epitopes, etc.). The design of the P-site™ epitope critically determines the outcome (yield, specificity and affinity) of the final antibody. Based on your individual project needs, our team will tailor the epitope design accordingly.
For a singly-phosphorylated peptide of 15 residues in length:
Yen-de-Zyn of P-site™ epitope
$195FREE
Synthesis of phosphorylated peptide:
HPLC high purity peptide will be used for immunization purposes.
General grade peptide for ELISA and Affinity Purification purposes.
Remaining amount of peptide will be provided to the investigator.
$650
($25 per extra residue when exceeding 15 residues, should epitope design call for it.)
Synthesis of non-phosphorylated peptide counterpart:
General grade peptide will be used for ELISA and affinity absorption purposes.
Remaining amount of peptide will be provided to the investigator.
$200
Carrier Protein Conjugations of:
HPLC-highly purified phosphorylated peptide to carrier protein (KLH or THY) for immunization,
HPLC-highly purified phosphorylated peptide to GIgG for ELISA, and
General grade non-phosphorylated peptide counterpart to GIgG for ELISA.
$295
P-site™ Regular™ Antibody production (2 Rabbits):
Regular™ Antibody Production regimen
~5 ml pre-immune serum per rabbit
~5 ml test bleed serum per rabbit
Pre-immune vs. Test Bleed ELISA against phosphorylated and non-phosphorylated peptides
~50 ml final antiserum per rabbit
$900
Step 1. Phosphorylated peptide-specific Affinity Purification of Antibody:
Preparation of phosphorylated-peptide conjugated affinity matrix
Separate purification of antiserum from both rabbits
Affinity matrix will be provided to the investigator upon completion of project
$845
Step 2. Affinity Absorption of Step 1-purified Antibody to separate P-site™-specific antibody from the cross-reactive antibody populations
Preparation of non-phosphorylated peptide conjugated affinity matrix
Separate absorption of both Step 1 purified antibodies
ELISA of pre- and post-purified serum to ensure depletion of specific antibody and ELISA of purified and absorbed antibody preparations against phosphorylated and non-phosphorylated peptides to ensure P-site™ specificity
SDS-PAGE of final antibody preparations
Affinity matrix will be provided to the investigator upon completion of project
$845
TOTAL
$3,735 $2,895*
*Pricing will be dependent upon type and/or number of modifications and epitope length.
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