Research Department
Analytical Core Facility
Director: Dr. Hans Peter Bächinger
503-221-3433
Manager: Dr. Kerry
Maddox
503-221-3445
The analyses performed by the
Analytical Core Facility rely on analytical instrumentation that is expertly
maintained and operated. In order to
provide analyses that are immediately useful and functional to users of the
facility, the staff provides data interpretation and assists in experimental
design. The core facility educates the
users in the capabilities of the instrumentation and keeps up-to-date with the
rapidly changing technologies. Along
with this, the staff develops methods that are needed for the successful
completion of research projects. The
high standards set by the Analytical Core Facility have a large impact on the
success of the research conducted at the Portland Unit.
The core facility consists of
three sections:
1. Protein Analysis Section provides sequencing of peptides and proteins,
synthesizes peptides, performs amino acid analysis and provides expertise in
protein biochemistry.
·
Peptide Synthesis
2. DNA Analysis Section sequences DNA and
synthesizes oligonucleotides and provides expertise
in DNA technologies.
·
DNA Oligo Synthesis
Request
3. Biophysical-chemical Analysis Section is responsible for
characterizing the hydrodynamic, thermodynamic and optical properties of
macromolecules.
Protein Analysis Services
Protein
sequence analysis.
The facility operates an ABI Procise Sequencer
for samples in solution or transferred to a PVDF membrane.
When submitting samples please provide
handling descriptions, buffers used, and amounts of protein. Samples in 10-20 pmole
range is necessary for optimal results.
Include a photograph of the gel if available.
Peptide Synthesizer
The facility uses an ABI 433A Peptide Synthesizer that was purchased in
2003 and maintained under service contract agreement.
Sample Preparation
The Facility has 2 Agilent 1090 HPLC systems
for purification and peptide separations.
These systems can be used as analytical instruments for peptide mapping
or as semi-preparative systems for other analyses. When internal sequence analysis is needed on
proteins, the samples are digested with a protease or chemically cleaved to
generate smaller peptides. These are
then separated on a choice of reverse phase, ion-exchange, or size-exclusion
columns and fractions collected.
Individual fractions are applied to the Procise
Sequencer for sequence determination.
Mass Spectrometry
The Facility purchased a Waters Micro-Q-Tof Mass Spectrometer in 2003 to
provide accurate mass determination.
This instrument is primarily used for screening synthetic peptides,
determining oxidative state of peptides, and peptide finger-printing for
protein identification. The HPLC component is a Waters Alliance 2695. MassLynx is the
operating and analysis software for the LC-MS.
Both instruments are covered under service contract agreements for
on-site service and technical support by telephone.
DNA Services
- DNA
sequencing
ABI 3100 Avant with 4 capillaries
- Oligonucleotide Synthesis
We are currently using an ABI 392
Synthesizer that contains 2 columns for dual synthesis.
Outsourcing DNA services:
When our sequencing capabilities become saturated, lab staff is
encouraged to send samples to the OHSU Core Facility (http://www.ohsu.edu/research/core/
).
For oligonucleotide synthesis requests that
cannot be met by our service, either for the volume of oligonucleotides
requested for special treatment, a number of companies are used off-site.