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CUSTOM ANTIBODY Against Post-Translational Modifications

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This service concern different modifications such as:

  • Phosphorylations
  • Methylations
  • Nitrations
  • Acetylations
  • Glycosylation
  • ....
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It consists in a classic Anti-peptide project wth some differences described as following. Please feel free to contact us, we will be happy to help you in your project.

 
  • Peptide synthesis

For this service, two peptides will be synthetizes:

    • One peptide without modification: <15 AA, 5-10 mg at 70% pure
    • One peptide with one modification: >15 AA, 20-25 mg at 70% pure
 
Nevertheless you can choose to synthetize peptides with otther quantities, purities, sequence length...
For more information, please contact us.
 
  • Conjugation
Usually we conjugate the peptide to a carrier protein via a cysteine residue. If the amino acid sequence of the peptide does not contain a natural cysteine, we add a cysteine to Nter or Cter sequence of the peptide.
    • Only the modified peptide is conjugated on carrier protein
    • Carrier proteins available: Keyhole Limpet Hemocyanin (KLH), Bovine Serum Albumin (BSA) or Ovalbumin (OVA)
    • Quantity of peptide conjugated: 5mg (exception when quantity conjugated producing antipeptide antibodies in goats : 10mg )

For this service, two rabbits will be immunized with the modified peptide according to our 89 days protocol.
This protocol is also totally adaptable. Then at the end of the standard protocol, you can request:
    • The final bleeding of animals
    • One or several intermediary bleeds in order to obtain a bigger volume of serum
    • To continue your immunizations in order to try to improve the seum's immuno-reactivity
 

Each sample will be tested for anti-peptide antibody using CovAbtest®

It enables the serum's immunoreactivity to be tested on each peptides, previously coupled covalantly to the wall of 96-microwell plats.
 
These elements and the results obtained on your lab enable you to take the right decision at the end your standard protocol.
 

This type of purfication will enable you to immunopurify the antibodies specific for the peptides and as a consequence to decrease background dering your experiments.
 

This service consist of:

    • Preparing one imunopurification column per peptide: 1mg of each peptide conjugated to 1ml of sepharose
    • Immunopurification in two step:
      • firstly, passage of 5ml of sera against the non-modified peptide sepharose column
      • secondly, passage of the flow through against the modified peptide sepharose column
    • Control of the immuno-reactivity of the antibodies immuno-purified by ELISA CovAbtest® against each peptide

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