*Synthesized peptides (≥10 aa) can be biotinylated to ensure better and oriented fixation onto the support.

Epitope mapping process

1. Peptide design

Peptides are synthetized in a 96-well format and can be biotinylated on request*.  You  provide the protein sequence, the desired peptide length (13 mer peptides are suggested) and overlap (7 aa overlaps are suggested), and we take care of generating the library.

Each 96-well plate is coated with 24 different peptides in triplicate.

Your Primary Antibody should be provided purified; A 1 mg/mL concentration in PBS is recommended. The mAb (dilution 1:1000) is added to each well, and a positive control Ab can be used (optional).

Detection is performed using an appropriate, HRP-labeled secondary antibody:

  • Anti-rabbit
  • Anti-rat
  • Anti-mouse
  • Anti-guinea pig
  • Anti-chicken
  • Anti-human or other on request

The positive peptides (triplicate) are incubated with various antibody dilutions (1:1000, 1:3000 and 1:9000).

Disclaimer : Because the strategy relies on synthetic, linear peptides, it may fail identifying the exact binding site of mAbs which target conformational epitopes.

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