Fastest Custom Monoclonal Antibody Services

Phospho-Specific Monoclonal Antibody Package Protocol

MonoExpress phospho-antibody package (SC1659)

Time Line
Phase I: Immunogen preparation
  • Peptide synthesis: 5 mg of phospho-peptide (up to 15 residues) with >90% purity and 3 mg non-phospho-peptide with >70% purity is synthesized by GenScript. 2 mg of phospho-peptide and 2 mg of non-phospho-peptide will be delivered to the customer along with the final antibody product.
  • Conjugation: The modified peptide is conjugated to the carrier protein KLH.
28 days
Phase II: Immunization and cell fusion with MonoExpress™ protocol
  • Immunization: Balb/c mice or rats are immunized and boosted with MonoExpress™ immunization protocol.
  • Fusion: Electrofusion is performed and at least 10,000 clones can be obtained from each fusion. Additional fusions are available to reach our guarantee without extra charge.
45 days
Phase III: Screening and supernatant samples preparation for 1st delivery
  • Screening and isotyping: The hybridoma cells are screened by ELISA with phospho and non-phospho peptides and all positive clones are expanded into 24-well plates. Up to 5 clones with the best ELISA value and specificity to phospho-peptide are selected for isotyping.
  • 1st delivery: 2ml supernatant samples for each clone are prepared for 1st delivery. Antibody Certificate of Analysis report is prepared with ELISA data and IgG concentration for each sample.
Phase IV: Subcloning, expansion, cryopreservation, antibody production and final delivery
  • Subcloning, expansion and cryopreservation: Up to 5 clones are subcloned by limiting dilution, expanded and frozen.
  • Antibody production: At least 2 mg purified antibody is produced for the clone with best screening result. The cross reactivity with non-phospho peptide < 10% for the purified antibody.
  • Final Delivery: Two vials of cell culture for each cell line, at least 2 mg qualified purified antibody, 2 mg of unconjugated phospho-peptide and 2 mg of non-phospho peptide are delivered to the customer.
 30 days

Note: GenScript provides free cell line storage service for 6 months from the date of cell freezing. If continued storage service is required after the end of six-month period, the charge is set at $10 per cell line per year.

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