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24 May 2017

Highly Specific Antibodies against Peroxiredoxin

Cambridge Research Biochemicals

Highly Specific Affinity Purified Polyclonal Antibodies

Circadian rhythms affect many metabolic pathways, for example the phylogenetically conserved peroxiredoxin (PRX) proteins, show daily rhythms in oxidation state. To explore the biochemical properties of the PRX system, Dr John O’Neill, based at the Laboratory of Molecular Biology in Cambridge enlisted the assistance of
Cambridge Research Biochemicals to prepare a custom made high affinity rabbit polyclonal anti-PRXSO2/3 antibody. The polyclonal anti-sera demonstrates a clear circadian rhythm in the over-oxidation of PRX in head lysates of Drosophila melanogaster sampled under constant dark conditions.

Right: Peroxiredoxin oxidation cycles in fruit flies. Two independently generated polyclonal antisera against over-oxidized peroxiredoxin  demonstrate circadian pattern in the overoxidation of PRX in head lysates of Drosophila melanogaster.

Above: This was sampled under constant dark (DD) conditions (the 2nd and 3rd DD cycles are shown). The custom-made antiserum was generated by Cambridge Research Biochemicals using a “double purification” approach: The crude serum was first depleted of non-sulfonylated epitopes using a peptide containing serine followed by retrieval of antigen-specific antibodies against the antigen peptide (sulfonylated sequence, DFTFV[C-SO3]PTEI).
CRB are experts in generating highly specific antibodies and have perfected such methods during their 37 years of expertise in the Life Science industry.

Publication reference: O’NEILL, J et al, Analysis of the Redox Oscillations in the Circadian Clockwork, Methods in Enzymology, 2015, 552, 185-210

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Target antibodies from CRB

CRB offer over 37 years of expertise in the generation of target specific polyclonal antibodies
against highly pure peptides made in-house by CRB‘s expert peptide chemists. Consultation on peptide design and perfected purification techniques help to guarantee project success and the project can be tailored to suit the required budget.
CRB also have an excellent publication record which can be viewed here.

TESTIMONIAL

DR JOHN O’NEILL – MRC LMB

John O'Neill MRC-LMB

“In many different model organisms and cell types, we had observed a circadian rhythm in the oxidation state of the highly conserved 2-cys peroxiredoxin family of antioxidants. As we investigated the underlying mechanism however, we found we were hindered by variations in the specificity of commercial available antisera against this oxidative protein modification.  CRB generated a highly specific affinity-purified polyclonal, additionally depleted for activity against unmodified peroxiredoxin.”

“This tool allowed us to further validate our original findings and obviated any need to take batch variation into account, thereby increasing experimental reproducibility. CRB’s service was efficient and professional, and very much tailored towards our particular needs. I do not hesitate to recommend them to colleagues.”

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