Phosphorylated Antibody Customized Service

Instruction manual

First Edition (Revised on April, 2016)

Post-translational modifications are essential for biological functions of proteins. In lots of crucial regulation mechanism of life, phosphorylation of proteins is an important post-translational modification, it is related to kinds of biological issues, such as signal transduction, cell cycle, growth and development, mechanism for cancer and etc.. Cloud-Clone Corp. provides phosphorylated polyclonal/monoclonal antibodies customized service. The ‘one-stop service’ from antigen design to antibody preparation is provided, which is quick and convenient. In the field of antibody production and development, Cloud-Clone Corp. produced a high polymer adjuvant, which shows better immune effect than traditional Freund’s adjuvant, and developed ‘Easy-fusion’ high efficient fusion technology. These contribute to the research and development of phosphorylated antibodies, which specifically target the phosphorylated protein with high sensitivity in the detection.
The typical process map of customzied phosphorylated antibody is shown as below,
Type I: Phosphopeptide — Animal Immune — Antiserum Collection — Antigen specific affinity Chromatography purification — Detection
Type II: Phosphopeptide — Animal Immune — Cell Fusion — Cell Screening —Ascites Preparation — Antibody purification — Detection
More information about customzied phosphorylated antibody.

Type	Materials Provided by Customers	Main Processes	Feature	Deliverable	Leading time
Type I: Preparation of Phosphorylated Polyclonal Antibody	Protein information	1. Polypeptide design 2. Polypeptide synthesis of both Phosphorylation & nonphosphorylated peptide 3. Polypeptide coupling 4. Physical examinations and serum collection for animals before immunization 5. Primary immunization with antigen 6. Blood collection, ELISA detection for serum samples 7. Secondary immunization 8. Blood collection, ELISA detection for serum samples 9. Third immunization 10. Blood collection, ELISA detection for serum samples 11. Forth immunization 12. Blood collection, ELISA detection for serum samples 13. Affinity purification and reverse screening 14. Qc test for cross-reactivity.	1. Serum collection before immunization,  ELISA detection for serum samples collected after each immunization 2. For primary immunization, Freund’s complete adjuvant or a polymer complete adjuvant with high efficient are provided for choose. 3. For the secondary, third and forth immunization, Freund’s incomplete adjuvant or a polymer incomplete  adjuvant  are provided for choose. 4. Affinity purification and reverse screening to increase specificity. 5. Purified Antibody titer for ELISA > 1:64,000, and cross-reactivity with nonphosphorylated polypeptide < 10%	1. Serum collected before immunization. Volume:1-2ml 2. Report for detection of serum samples 3. Purified Antibodies:1-10mg 4. Detection report for antibodies 5. Remaining polypeptide 6. Report of polypeptide synthesis and modification	12-20 weeks
Type II: Preparation of Phosphorylated Monoclonal Antibody	Protein information	1. Polypeptide design 2.Polypeptide synthesis of both Phosphorylation & nonphosphorylated peptide 3.Polypeptide coupling 4.Physical examinations and serum collection for animals before immunization 5.Immunization of animals 6. Cell fusion and screening 7. Preparation of ascites 9. Purification off antibody 10. Qc test for cross-reactivity.	1. Cloud-Clone Corp. provides multiple  carriers for selection 2. The Nano emulsion technology and a polymer adjuvant  can improve the animal immunization effect 3. Five Balb/c mice or three rats are offered 4. ‘Easy-fusion’ high efficient fusion technology, rapid and convenient, high rate of fusion and success 5. A wide variety of choices for antibody purification available 6. Purified Antibody titer for ELISA >1:64,000, Cross reactivity with nonphosphorylated polypeptide<10%.	1. 1~3 hybridoma cells 2. ≥1mg antibody (Select one strain to produce antibody) 3. Detection report for antibodies 4. Remaining polypeptide 5. Report of polypeptide synthesis and modification	20-25weeks