The Protein Nucleic Acid Chemistry Laboratory
The Protein Nucleic Acid Chemistry Laboratory offers advanced analytical technologies to the University of Leicester, UK and international academic institutions and other external clients.
PNACL has been providing excellence in analytical services and collaborative research since 1996. The laboratory is headed by Dr Sharad Mistry with expertise in proteomics and mass spectrometry provided by Dr Andrew Bottrill. Analysis type falls into two broad categories:
Services for full template preparation through to simple low-cost analysis of completed sequence reactions are available. The facility is equipped with a high-throughput Applied Biosystems 3730 Genetic Analyser for running up to 48 samples concurrently. In addition, samples can be submitted for DNA fragment analysis for applications such as microsatellite analysis or Amplified Length Polymorphism (AFLP) analysis. Big Dye and a number of common sequencing primers (including 47 PROTEX pLEICS primers) are kept in stock.
Facilities for carrying out protein identification, quantification and determination of post-translation modification are available. These include mass spectrometry platforms, enabling high resolution analysis utilising an LTQ-Orbitrap instrument and high sensitivity diagnostic validation using a 4000Q-Trap mass spectrometer. Advanced data analysis using a variety of proteomic software platforms can be carried out.
Pure proteins (from gel or in-solution) can be identified by trypsin digestion followed by MALDI-ToF mass spectrometry and database searching.
LC-MS/MS can be used to identify multiple proteins and sites of post-translational modifications. For some samples additional fractionation or modification specific enrichment may be required, and these facilities are available.
Protein Quantification. Subtle changes in protein abundance can be detected using a variety of labeling techniques (SILAC, iTRAQ, TMT) or via label free approaches (Spectral Counting, SRM).
Protein Molecular Weight can be determined by either Electrospray mass spectrometry or MALDI-ToF MS.