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.

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

“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.”