Custom Polyclonal Antibody Production Service
Phase 1. Peptide synthesis
Custom FMOC Peptide synthesis up to 20 amino acids (20 mg). 10 mg of free peptide will be delivered to the customer together with the antibody.
A cysteine will be automatically added to the C-terminus or N-terminus of the peptide, and conjugated to KLH
Assistance on designing peptide with respect to protein homology, antigenicity, hydrophilicity, and synthetic suitability.
Peptide purity is >80%.
Time: 2 weeks
Phase 2. Peptide conjugation to KLH
Conjugation of 10 mg peptide (using Cysteine) to KLH.
Time: 3 days
Phase 3a. Immunization and anti-sera production in rabbit
Conjugated peptides or recombinant proteins are used to immunize rabbits.
Antigen provided by the customer: 10 mg of peptide or protein, the concentration should be >0.4 mg/ml and the buffer cannot contain organic solvents.
Ten-week protocol to produce antibodies in 2 SPF rabbits.
ELISA tests until the titer reaches 1:4,000.
Western blot analysis when ELISA reaches target titer.
3ml of pre-immune serum and 60ml of final immune bleeds.
Time: 8-10 weeks
Phase 3b. Immunization and anti-sera production in mouse
Conjugated peptides or recombinant proteins are used to immunize mice.
Antigen provided by the customer: 5 mg of peptide or protein, the concentration should be >0.4 mg/ml and the buffer cannot contain organic solvents.
Ten-week protocol to produce antibodies in 5 SPF mice.
ELISA tests until the titer reaches 1:4,000.
Western blot analysis when ELISA reaches target titer.
0.1ml of pre-immune serum and 3ml of final immune bleeds.
Time: 8-10 weeks
Phase 4. Antibody purification
Purification of the antibody by protein G.
Antibodies are eluted using both pH11 and pH2.5 buffers and neutralized immediately followed by dialysis in PBS.
Time: 1 week
