Protein Technologies Symphony and Tribute Peptide Synthesizers. The Tribute™ is a 2 channel bench top peptide synthesizer with a simple design and setup. The Symphony® is a 12-channel synthesizer where each reaction channel operates independently of the others; analogous to having 12 single channel machines. The benefits and advantages of t he Symphony® include versatility, fast multiplex operation, low coupling cost, variable synthetic scales, automated cleavage, and customizable protocols

AKTAExplorer FPLC system (Amersham/Pharmacia): The laboratory has a fully automated chromatographic system AKTAExplorer FPLC system (Amersham/Pharmacia). ÄKTAFPLC is a preparative liquid chromatography system for fast and easy purification of proteins. The entire system is refrigerated and computer controlled using Unicorn software v3.10.

Profinia™ Affinity Chromatography Protein Purification System. The system performs automated two-step affinity purification. It replaces traditional gravity-flow techniques with a convenient, complete system for the purification and desalting of affinity-tagged proteins. Histidine (His)-tagged, GST-tagged, affinity, and desalting/buffer exchange methods deliver purity and yield comparable to other techniques, and require only a fraction of the time to obtain protein ready for experiments. Profinia software offers a convenient way to review purification results. Its user interface is easy to use yet comprehensive and customizable for either real-time data acquisition or data analysis.

Protein production, purification and characterization: PCL primarily utilizes E. coli, insect cells (baculovirus-mediated) and mammalian cell expression (transient expression, and virus-based expression of proteins). In addition, PCL can construct cell lines permanently expressing a gene of interest. PCL can undertake specialized projects in collaboration with CEIDR GeneLab and the Molecular Immunopathology & Microscopy Core (MIPC).

Expression of proteins in insect cells: PCL specializes in the production of proteins using the baculovirus-based expression systems in conjunction with the Sf9, SF21 and Hi-five insect cells. Recombinant baculoviruses are constructed using commercially available, as well as modified baculoviruses, in collaboration with the CEIDR/GeneLab Core. Insertion of appropriate PCR product is confirmed, using DNA sequencing and, expression of desired proteins in small scale insect cell cultures, using indirect immunofluorescence and western immunoblotting assays. Small-scale test expressions in both suspension and adherent cultures are conducted to determine optimal expression conditions prior to scaling up to larger cultures in roller bottles, or bag-based cultures. The Core has the capacity to carry out up to 20 roller bottles in specialized incubators, or 18L cultures at a time using shaking suspension culture expression runs.

Mammalian Cell Expression experiments: The Protein Core Laboratory maintains Chinese hamster ovary (CHO-S; free-style) cell lines cultured in minimal media. Also, additional cell lines are available including HEK 293-F (free-style), as well as conventional cell lines including African green monkey (Vero), and other cell lines primarily devoted for transient expression experiments. The Core can undertake the construction of stable, clonal cell lines expressing proteins of interest as specialized collaborative projects or as fee-for-service. Typically, these expression experiments are carried out in roller bottle apparatus within specialized incubators or using bag-based cell culture systems. Maximum capacity for suspension cultures is approximately 18L for either suspension or adherent cultures.

The Protein Laboratory is home to our Peptide, antibody, production and protein interaction studies facility. This laboratory concentrates on the production and purification of proteins and antibodies under contractual arrangements with clients. Towards this end we have at our disposal a wide array of instruments ranging from the Protein Technologies Symphony and Tribute Peptide Synthesizers, The affinity based Profinia and AKTA explorer protein purification systems, the Blitz and the Biacore X100 for studying protein protein interactions. The laboratory also produces monospecific antibodies in rabbits and mice using conventional immunization methodologies using purified protein immunogens as well as genetic immunization methods. Antibodies are concentrated and purified using standard methodologies. The laboratory provides service support on the use of chromatographic methods for the purification of proteins from bacterial or eukaryotic expression systems. One example of a purification procedure involves the purification of histidine-tagged proteins produced in bacteria and insect cells. Purification by FPLC is done using immobilized metal ion adsorption chromatography (IMAC).

The Protein Laboratory is home to our Peptide, antibody, production and protein interaction studies facility. This laboratory concentrates on the production and purification of proteins and antibodies under contractual arrangements with clients. Towards this end we have at our disposal a wide array of instruments ranging from the Protein Technologies Symphony and Tribute Peptide Synthesizers, The affinity based Profinia and AKTA explorer protein purification systems, the Blitz and the Biacore X100 for studying protein protein interactions. The laboratory also produces monospecific antibodies in rabbits and mice using conventional immunization methodologies using purified protein immunogens as well as genetic immunization methods. Antibodies are concentrated and purified using standard methodologies. The laboratory provides service support on the use of chromatographic methods for the purification of proteins from bacterial or eukaryotic expression systems. One example of a purification procedure involves the purification of histidine-tagged proteins produced in bacteria and insect cells. Purification by FPLC is done using immobilized metal ion adsorption chromatography (IMAC).

The laboratory also produces monospecific antibodies in rabbits and mice using conventional immunization methodologies using purified protein immunogens as well as genetic immunization methods. Antibodies are concentrated and purified using standard methodologies. The laboratory provides service support on the use of chromatographic methods for the purification of proteins from bacterial or eukaryotic expression systems. One example of a purification procedure involves the purification of histidine-tagged proteins produced in bacteria and insect cells. Purification by FPLC is done using immobilized metal ion adsorption chromatography (IMAC).

Glen-Mills French Press High pressure Homogenizer. For disruption of cellular plasma membranes / internal organelles / homogenzation of animal tissue.

Fortebio Blitz Protein Assay system. Blitz™ is a personal assay system, letting you do label-free analysis. The user can easily test column fractions for presence of protein of interest, check protein activity and quantitate before starting a complex experiment, characterize proteins in cell lysates in their folded state or monitor protein expression. BLItz gives results in a just a few seconds or minutes using just a drop of the sample.