Protein Purification
Chromatography Systems
- Pharmacia FPLC System
- Low-pressure chromatography
- Columns for size-exclusion, anion and cation exchange
- Can be run in the cold to preserve biological activity of compound of interest
- Please contact the facility if you are interested in this service and for training on the use of the instrument (806)742-3722 X 225
- More Information.
- Applied Biosystems BioCad Sprint
- Medium-pressure, rapid chromatography
- Monitor two wavelengths simultaneously (uv or visible)
- Please contact the facility if you are interested in this service and for training on the use of the instrument (806) 742-3722 X 225
- More Information.
Electrophoresis Systems
- Biorad Protean IEF 2D-gel System
- First dimension: isoelectric focusing of proteins
- Second dimension: protein separation on SDS-Page
- In-gel digestion of protein spots analyzed
- Please contact the facility if you are interested in this service and for training on the use of the instrument (806) 742-3722 X 225
- More Information.
Pharmacia FPLC Information
This column chromatography system automates the separation and purification of semi-preparative amounts of the compound of interest, typically a protein. The system uses aqueous buffers and fairly low pressures, and is housed in a "cold box" so that chromatographic runs can be done at 4°C, thus helping to preserve biological activity. The two pumps allow for gradient elutions (for example, increasing salt concentration on an ion exchange column), and the fraction collector permits automated collection of fractions from the column by volume, time or peak detection. There are currently a choice of five different columns: Superdex 75 and Superose 12 for gel filtration (fractionation range approximately 3-70FD and 1-300KD respectively); Mono Q for anion exchanges; mono S for cation exchange; and phenyl sepharose, a hydrophobic interaction matrix. Users must supply their own buffers. Samples or buffers containing detergents can only be used on columns which have been so designated.
Training on the instrument and fees to cover the cost of consumables and column use will be determined during discussion of the separation/purification project.
Applied Biosystems BioCad Sprint Information
This column chromatography system (formerly made by Perseptive BioSystems) is capable of performing separations on POROS ion exchange columns in as little as 5 minutes at a semi-preparative level.
The instrument can monitor two wavelengths (either UV or visible) simultaneously and has in-line conductivity and pH monitors which permit the precise monitoring of salt and pH gradients during the chromatographic run. There are adapters to allow use of the FPLC columns mentioned in the section above.
Training on the instrument and fees to cover the cost of consumables and column use will be determined during discussion of the separation/purification project.
Biorad Protean IEF 2D-gel Information
The newest addition to the core facility is the BioRad 2-dimensional gel electrophoresis system. 2D-gel electrophoresis is a powerful technique for separating and isolating a complex mixture of proteins. The first dimension separates the proteins on the basis of their isoelectric point. These proteins are separated further in the second dimension on the basis of their size (standard SDS-PAGE). Individual spots (proteins) from the stained gel can be selected and subjected to in-gel proteolysis and peptide extraction. The extracted peptides can be analyzed by mass spectrometry for identification.
Three size formats are available, depending on the complexity of the protein sample. The first dimension (isoelectric focusing) is run on the Protean system, with a selection of pH ranges and IPG strip lengths available. The second dimension (SDS-PAGE) may be run using the Protean IIxi cell (for 17cm gels), the Criterion cell (for 11cm gels) or the Tetra cell (for 7cm gels). Please consult with the Center staff regarding the use of this system for your research goals.