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-: My name is Sophie Alvarez
and I'm the director of the Proteomics
and Metabolomics Facility
which is part of the Nebraska center for biotechnology.
Our facility is located on the first floor
of the Beadle Center.
The facility was established back in 2015
to meet an ever-increasing need
for a range of technical services using mass spectrometry.
These services include identification
and relative quantification of proteomics
and several advanced methods for profiling
and quantitation of small molecules.
The primary aim of the facility
is to provide high quality service and expertise
for faculty, students, and staff at UNL
in the fields of proteomics and metabolomics.
To accomplish this goal, the facility is equipped
with state of the art instruments.
The proteomics platform includes an Orbitrap Eclipse Tribrid
Mass Spectrometer coupled to a Dionex U3000 Nano HPLC
to provide extremely high resolution,
high mass accuracy data.
We routinely run comprehensive experiments
for protein identification and differential proteomics
using either label-free or labeling approaches.
The labeling method utilizes the tandem mass tag
or TMT chemistry, which allows to combine up to 16 samples
into one run, reducing experimental variability.
The total number of protein identified
ranges from 2,000 to 8,000 proteins
depending on the experiment design.
This platform is also used to identify proteins
from cool-down acids in interactions to these
of single proteins from purification experiments.
For metabolomics, these needs are addressed
by the use of multiple instrument capabilities.
For more basic analysis we use a standalone
Agilent Ultra-.High Performance Liquid Chromatography System
or UHPLC which is equipped with a diode-array detector,
a fluorescence detector and evaporative light
scattering detector or yellow steel.
This provides a robust cost effective platform
to establish targeted assay of a wide range
of compounds from common molecules like amino acids
and carbohydrates to more specialized metabolites
like carotenoid and phospholipids.
Our gas chromatography mass spectrometer or GCMS
allows for the detection
of volatile compounds found in foods
of all the tissue samples and offers a targeted approach
to studying polar metabolites,
such as those involving carbon metabolism.
For projects requiring increased sensitivity
when compounds are a very low abundance,
the facility offers a liquid chromatography MS platform.
This consists of a 6,500 Qtrap mass spectrometer
with a Shimadzu UHPLC on the front.
With this we can very carefully target particular molecules
using a technique called multiple reaction monitoring
or MRM for short, example of the MRM assays available
on this platform include phytohormones, flavonoids,
alpha and beta acids from hops and by acids.
All of these three pieces of equipment, the HPLC, GCMS
and the Qtrap are open access for students, staff
or faculty to set up their own methodologies
and training to do this is provided by the facility.
Finally for LCMS and targeted metabolomics
we offer our Q Exactive High Field Orbitrap or QEHF.
This is a high resolution accurate mass system
coupled to a thermal vanquish UPLC.
It is set up to run reverse space
and hydrophilic interaction liquid chromatography
for increased metabolomic coverage.
For all the platforms mentioned both proteomic
and metabolomics the post acquisition data analysis
is supported by several established pipelines
of bioinformatic tools run on powerful computing servers
and workstations, with these specialized platforms
the core facilities brought impact
on research includes, advancing world-class research,
supporting grant proposals, assisting
in the education of future scientists
and supporting economic and scientific advancements.
Our outreach and education program includes workshops
seminars, and courses.
We also provide the services to academic labs outside
of Nebraska and to private industry.
Full service and self-service options
are able to meet the need and budget of our users.
For more information please visit our webpage
or follow us on Twitter.
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