The Cornell University Protein Facility (CUPF) was founded in 2000 with funds from the Tri-Institutional Research Program provided by The Atlantic Philanthropies, and the National Institutes of Health. As of 2006, the Facility is in the process of becoming a Cornell University service facility as a component of the Life Sciences Shared Core Facilities.
The objective of the Protein Facility is to provide investigators with equipment, expertise and custom services for the production and characterization of biomolecules.
The Protein Facility encompasses four major efforts:
- molecular biology (cloning, mutagenesis, etc.)
- prokaryotic protein production/purification (E. coli)
- eukaryotic protein production/purification (yeast, insect and mammalian cells)
- characterization instrumentation (binding studies, kinetics)
Each of these resources provides unique, cost-effective services and expertise to investigators. These services allow some groups to increase throughput by freeing them from repetitive protein purifications and standard molecular biology. For other groups the custom services allow expansion into new areas of investigation without a large investment of time, effort and training. The instrumentation resource allows investigators access to state-of-the-art equipment, having to make the financial commitment of a purchase.
The facility offers services and equipment access on a fee-for-service basis and confidentiality is maintained at all times.
Molecular Biology: Using standard techniques, the Protein Facility will clone genes from diverse sources (plasmids, genomic DNA, cDNA, libraries, etc.) into vectors for recombinant expression in various host systems (E. coli, yeast, baculovirus, mammalian, etc.). Additionally, we can create site-directed, deletion, insertion or silent mutants. Other molecular biology techniques are also available.
Prokaryotic Protein Production: The Protein Facility can produce protein in E. coli using a variety of expression vectors and conditions. Produced proteins can be purified using a variety of affinity techniques (if tags are available on the recombinant protein) or standard chromatography. We can scout conditions and fusion tags to attempt to find suitable conditions for any particular protein.
Eukaryotic Protein Production: The Protein Facility can produce proteins in yeast (S. cerevisiae, others), insect cells (using the baculovirus expression system) or in cultured mammalian cells (this service will begin in the fall of 2007). Once produced the protein can be purified as described for proteins derived from prokaryotic expression.
Instrumentation: The facility maintains a suite of instrumentation for the characterization of macromolecules and enzymatic reactions: an SF-2004 stopped-flow spectrophometer with a titration module (KinTek), an RQF-3 rapid quench flow instrument with freeze quench adaptor (KinTek), a Nano-ITC III isothermal titration calorimeter (Calorimetry Sciences Corporation), a Synergy HT Multi-Detection plate reader (BioTek), and an XL-I analytical ultracentrifuge (Beckman Coulter). Protein production instrumentation includes: three large capacity shakers (New Brunswick, LabLine), two incubators at 30°C and 37°C (VWR, Fisher), an Avanti J20-XPi centrifuge (Beckman Coulter), an Allegra 25R centrifuge (Beckman Coulter). The Protein Facility has equipment for cell lysis: Sonifier 250 sonicator (Branson), BioNeb (Glas-Col), Basic Z cell disrupter (Constant Systems) and a Emulsiflex C3 high pressure homogenizer (Avestin). Protein purification equipment includes: AKTA FPLC (GE Healthcare), AKTAXpress (GE Healthcare), Rotofor solution isoelectric focusing unit (Bio-Rad), Model 491 prep cell (Bio-Rad), and a variety of low-flow instrumentation (peristaltic pumps, fraction collectors, etc.), as well as a full range of columns for affinity or standard chromatography. The facility is fully equipped with all instrumentation necessary for performing DNA manipulations: plate format gradient thermocycler (Eppendorf), Storm phosphoimager (GE Healthcare), multiple agarose gel electrophoresis units, multiple SDS-PAGE gel electrophoresis units, vortexers, mini-shakers, microcentrifuges, etc. The facility has a tissue culture room equipped with CO2 incubators and a six foot BSL2 hood.
The facility is located in the ST Olin Chemistry Research Tower, rooms B77 and B78.