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Tip: How to detect small peptide clearly and sensitively by Western blotting or SDS-PAGE?
Peptide Antigen Design for Antibodies
peptide analysis tool for peptide antibodies
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LifeTein has developed the SIMBATM (SIngle Monoclonal B cell Antibody) technology based on single B cell retrieval to identify antigen-specific mAbs. The SIMBATM technology includes single cell RT-PCR with successional cloning and expression in vitro and requires relatively fewer cells. The SIMBATM technology is an attractive approach for retrieval of naive and antigen-experienced antibody repertoires generated in vivo and design of rationale structure-based vaccine.

A: (Cat. No. LA005) Monoclonal antibody services from gene to ascites Antibodies
  1. Gene amplification, vector construction, protein expression in E. coli., immunization and sera collection, ELISA test
  2. Splenocyte fusions are performed from mice with the best titers; The hybridoma cells are screened by ELISA
  3. Standard 22-26-week antibody production schedule
  4. Two of these 10 clones are selected for subcloning, isotype determination, ascites production and purification
  5. Deliverables: construct vector, 1 mg protein, 50 ul pre-immune sera (1:10 dilution), 2 clones, 2-5 mg purified antibodies (2 clones), ELISA results >1:32,000, isotype determination
B: (Cat. No. LA006) Monoclonal antibody services from peptide design to ascites production
  1. Include peptide design (free) and peptide synthesis, immunization and sera collection, ELISA test.
  2. Splenocyte fusions and subclone, ascites production and purification.
  3. Standard 20-26-week antibody production schedule
  4. Deliverables: 2-3 mg peptides, 50 ul pre-immune sera, 2 clones, 2-5 mg purified antibodies (2 clones), ELISA results >1:32,000, isotype determination
C: (Cat. No. LA007) Monoclonal antibody services using phospho-peptide antigens
  1. Peptide antigen design (free) and peptide synthesis
    • Phosphopeptide: >85% purity, 10 mg, 12-22 amino acids
    • Non-phosphopeptide: >85% purity, 10 mg, 12-22 amino acids
  2. Conjugation of phosphopeptide to KLH or BSA (2-3 mg); conjugation of the non-phosphorylated peptide to BSA; immunization and sera collection; ELISA identification; fusion and hybridoma development; isotype determination; subcloning and cell cryopreservation; ascites production and purification (2 clones)
  3. Standard 21-26 week antibody production schedule
  4. Deliverables: 50 ul pre-immune sera, 2-3 mg peptides (unmodified and phospho), 2-5 mg phospho-specific antibodies (2 clones), ELISA results >1:32,000, isotype determination
D: 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 25ml 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 request
  • Western blot: Assay of antisera or purified antibodies against the antigens, upon request
  • Immunochemistry: Assay of purified antibodies against the pathological slices, upon request

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Holocentromeres can consist of merely a few megabase-sized satellite arrays Nature CommunicationsNat Commun 14, 3502 (2023).

LifeTein designed and synthesized a series of peptides: CENH3, MIS12 and NDC80. The peptides were used for immunization of rabbits to generate polyclonal antibodies.

... The peptide synthesis, immunization, and antibody purification were performed by LifeTein...

Richard R.Rustandi, et al. Characterization of gH/gL/pUL128-131 pentameric complex, gH/gL/gO trimeric complex, gB and gM/gN glycoproteins in a human cytomegalovirus using automated capillary western blots; Vaccine; Available online 3 July 2021,

LifeTein generated Anti-gM and anti-gN antibodies for Vaccine Analytical Research Development and Vaccine Process Development Merck & Co., Inc., Kenilworth, NJ, USA. The rabbit polyclonal anti-gM and anti-gN antibodies were elicited by vaccinating animals with synthesized peptide sequences of gM (1-13aa and 345-372aa) and gN (61-101aa). Animal immunization and sera collection were performed at Lifetein, LLC (Hillsborough, NJ, USA). The sera were affinity-purified by its corresponding peptides.

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

George R. Tiller, Potential Roles of Peroxidases in Caenorhabditis Elegans Innate Immunity. 2014

Custom peptide antibodies were provided by LifeTein to study SKPO-1. The SKPO-1 peroxidase domain peptide was synthesized by LifeTein. The polyclonal antibodies were raised in rabbits and then affinity purified to accomplish high specifications and concentrations. The antibodies were successfully used to localize the SKPO-1 protein in hypodermis. It was found that the peroxidase SKPO-1 contributes to the host immune response during infection.

... With respect to having a pAb raised for IHC, I designed a synthetic SKPO-1 peroxidase domain peptide to be chemically synthesized and have a pAb raised in rabbits against said peptide by Lifetein®. This endeavor was successful and is covered in Chapter 3...

Hui-Lung Sun. et al. Stabilization of ERK-Phosphorylated METTL3 by USP5 Increases m6 A Methylation; Mol Cell; 2020 Nov 19;80(4):633-647.e7. doi: 10.1016/j.molcel.2020.10.026.

LifeTein helped synthesized a series of phosphorylated peptides. Then the peptides were used for phospho-specific p-METTL3 (S43) antibody productions. Sun et al. demonstrate that activation of the ERK pathway promotes m6A methylation by phosphorylation of METTL3 and WTAP.

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.

ERK Activation Globally Downregulates miRNAs through Phosphorylating Exportin-5. Cancer Cell. 2016 Nov 14;30(5):723-736. doi: 10.1016/j.ccell.2016.10.001.

LifeTein designed and synthesized a series of phosphorylated peptides. ELISA and dot blot analysis were performed for the verification of antibody specificity.

... Antibodies to the Thr345, Ser416, and Ser497 phosphorylation sites of XPO5 were generated in collaboration with Lifetein LLC... Supplementary information