Custom Polyclonal Antibody Services

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A: (Cat. No. LA001) Standard polyclonal antibody services using peptide antigens

1. Peptide antigen design (free): Let our highly skilled staff assist you with your antigenic selection and design. Simply send your protein sequences to us!
2. Peptide synthesis: >75% purity, 10 mg, up to 15 amino acids
3. Conjugation of peptide to KLH (2-3 mg)
4. Standard 12-15 week antibody production schedule including 4 immunizations with 2 rabbits
5. Affinity purification of selected batch of serum. Deliverables: 2-3 mg peptides, 1-2 ml pre-immune sera, 2-5 mg purified antibodies, ELISA results (>1:32,000)

B: (Cat. No. LA002) Polyclonal antibody services from gene to antibodies

1. Gene amplification, vector construction, protein expression in E. coli.
2. Immunization, antigen affinity purification
3. Standard 14-17 week antibody production schedule
4. Deliverables: construct vector, 1 mg protein, 1-2 ml pre-immune sera, 2-5 mg purified antibodies, ELISA results (>1:32,000)

C: (Cat. No. LA003) Standard polyclonal antibody services using phospho-peptide antigens. More on phosphorylation.

1. Peptide antigen design (free) and peptide synthesis
   • Phosphopeptide: >95% purity, 10 mg, up to 15 amino acids
   • Non-phosphopeptide: >75% purity, 10 mg, up to 15 amino acids
2. Conjugation of phosphopeptide to KLH or BSA (2-3 mg); conjugation of the non-phosphorylated peptide to BSA
3. Standard 13-16 week antibody production schedule including 4 immunizations with 2 rabbits
4. Affinity purification of selected batch of serum; depletion of non-phosphospecific antibodies; purification of phosphospecific antibodies
5. Deliverables: 1 pre-immune serum collection, 2-3 mg peptides (unmodified and phospho), 1-2ml pre-immune sera, 2-5 mg phospho-specific antibodies, ELISA results (>1:32,000)

D: (Cat. No. LA004) Protein Immunogens for polyclonal antibodies

1. Protein provided by customer (>5mg, >90% purity, no glycerol)
2. Protein characterization
3. Immunization and sera collection
4. Standard 15-18 week antibody production schedule
5. Deliverables: 0.5-1 ml pre-immune sera per rabbit, 60-80 ml antisera or protein A/G purified antibodies, ELISA results >1:12,000

Reference:

Natural variant of collagen-like protein A in serotype M3 group A Streptococcus increases adherence and decreases invasive potential. Infect Immun. 2015 Jan 5. pii: IAI.02860-14.

LifeTein made the affinity-purified rabbit polyclonal anti-SclA antibody. The purified streptococcal collagen-like protein A (SclA) antibodies demonstrated that a mutation in Group A Streptococcus carrier strains increase adherence and decrease virulence.

E: Additional services:

• 4-Week extension: We provide 1 additional immunization and 2 additional bleed collections for approximately 80-100ml of polyclonal antisera
• Affinity purification: Affinity purification of 25 ml of sera, including the affinity column for running additional purifications
• Antibody Conjugate Service: We can conjugate your antibody or protein to many labels: Biotin, Strepatvidin, HRP, AP, and fluorescent dyes.
• ELISA: On any bleed or purified antibody, upon requests
• Western blot: Assay of antisera or purified antibodies against the antigens, upon request
• Immunochemistry: Assay of purified antibodies against pathological slices, upon request

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Reference:

Jia Shen. et al. EGFR modulates microRNA maturation in response to hypoxia through phosphorylation of AGO2. Nature 497, 383–387 (16 May 2013), doi:10.1038/nature12080

LifeTein helped designed and synthesized a series of phosphorylated peptides. Then the peptides were used for phospho-specific antibody productions. The phospo-specific antibodies by LifeTein were confirmed to react with the epidermal growth factor receptor (EGFR). The Hung's lab showed that AGO2-Y393 phosphorylation mediates EGFR-enhanced cell survival and invasiveness under hypoxia. These findings suggest that modulation of miRNA biogenesis is important for stress response in tumour cells.

... The following peptides were chemically synthesized for antibody production in mice (Lifetein Conc.), Elisa verification (LifeteinConc.) and peptide competition assay in immunohistochemistry (IHC)... Supplementary information