Phospho-specific peptide design, peptide synthesis, immunization, affinity purification with affinity absorption - 91246PX
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Proteintech offers peptide synthesis containing a phosphorylated amino acid, conjugation to KLH, immunization of two rabbits, and affinity purification with cross absorption (Catalog# 91246PX). In the case a customer needs an antibody specific to the phosphorylated peptide that is non-cross-reacting with an unphosphorylated one, affinity purification with the phospho-peptide column, in addition to, cross absorption of the antibody with a non-phospho-peptide is required. Proteintech will offer cross absorption free of charge to save the customer the extra $399 (Catalog# 90006) for another round of purification.
The conjugated peptide will be used to immunize two rabbits. The customer can choose from one of the standard protocols for immunization. See Catalog#90001 for protocols. Proteintech will prepare an affinity purification column and purify 30ml of antiserum with the column then perform cross absorption. The customer will receive the extra unconjugated phospho- and non-phospho-peptides except for a small amount used for ELISA for the titer assay, the affinity column, purified anti-serum, flow through, and the rest of the anti-serum from the two rabbits. If the customer would like any additional phosphorylated amino acids, it will cost $150 each.
The service also includes the following:
1) Free phospho-peptide Design: Upon customer request, the technical support team of Proteintech will review the protein sequences and rank the most antigenic domains and submit them to the customer for a decision.
2) A large amount and high purity phospho-peptide synthesis: The chemists at Proteintech will synthesize the phosphor-peptide ordered. The yield of synthesis will be approximately 40 mg, at approximately 90% purity for 20 mer-peptides with Mass Spectrum data. There will be no need for additional purification, therefore you do not have to pay for the purification cost, which is a significant savings. We will charge $20 for each additional amino acid.
3) A large amount of KLH conjugation: 10 mg of phosphor-peptide will be conjugated with 5 mg of KLH protein (Catalog# 90004).
90001................Immunization (rabbit polyclonal antibody production)
90004................Peptide conjugation to KLH
90124.................Peptide design, peptide synthesis and immunization
90124P...............Phospho-specific peptide design, peptide synthesis and immunization
91246P...............Phospho-specific peptide design, peptide synthesis, immunization and affinity purification
Which Protocol should I use?
The standard long protocol is the preferred protocol because it has been widely used and tested through history. The long protocol provides a better success rate than the short protocol. The short protocol is designed for customers who need the antibody in a rush and are willing to trade success rate for time.
How much antiserum will I receive?
Each rabbit will give ~12 ml of serum in each production bleed and 45-50 ml of serum (from 90 - 100 ml blood) in the final bleed, a total of about 150 ml of antiserum from two rabbits
How does Proteintech determine which rabbit’s serum to purify?
We decide that by the ELISA result with test bleed. If both rabbits give good results, we will purify 15ml of serum from each rabbit. If one rabbit gives a better result, we will purify 30ml of serum from this "good" rabbit.
How much purified antibody will I receive?
It depends on the concentration of the specific antibody in the serum provided. Usually we can get 2-10ml from 15ml of antiserum.
- How do I order this service?
Download and print out a custom order form, fill out the relevant information, and fax to 312-455-8408 (USA and unlisted regions) or (+44) 161 23 21 272 (Europe). Once we have received your forms someone will contact you to discuss further your order and direct you where to send your protein sample.
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