Download P F I

I MAGE G ALLERY
Probing 100 fmole of digested HSA by Nano LC-LTQ/MS/MS
P ROTEOMICS F ACILITY
S CIENTIFIC O BJECTIVES
Proteomics Facility at the University of Vermont is
a core facility funded by the Vermont Genetics
Network to serve the Lead Institution and research throughout the state as part of the IDeA
Program mission of building research infrastructure. The facility provides a central resource of
mass spectrometry-based proteomics technologies
to identify, characterize and quantify target proteins in various biological and biomedical samples. We strive to establish a highly efficient research and educational environment for sharing
ideas, experiences, and knowledge of proteomics
application in systems biology, biomedical and
clinical studies.
UVM/VGN
PROTEOMICS FACILITY
High Quality for Low Rates!
Determination of Phosphorylation Sites in GSK3-beta after Drug Treatment
Consultation
Free
Coomassie Blue-stained gel band
$10/band
Silver-stained gel band
$25/band
Data analysis (bioinformatics)
Free
Method development
Free
Marsh Life Science 335
F ACILITY
Number of sample submissions
I NFORMATION
Number of facility users
VGN Proteomics
University of Vermont
Marsh Life Science 335
Burlington,VT 05405
Identification of Pafa1b2 from Human Testis Homogenate
http://vgn.uvm.edu/proteomics
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Quantification of Protein Expression in Biomedical Samples
More information about our facility can be found
on our website (http://vgn.uvm.edu/proteomics)
VGN is funded by a grant from the National
Institute of Health
F ACILITY I NFORMATION
Services Provided:

Protein identification, isoform characterization and
molecular weight determination

Identification of posttranslational modifications (PTMs;
e.g. phosphorylation, N-glycosylation, methylation,
acetylation, ubiquitination, tyrosine nitration,
S-nitrosylation, etc)

Identification of protein-protein interacting partners and
determination of their binding stoichiometries

De novo sequencing of novel proteins

Mass spectrometry (MS)-based global quantification of
protein expression and PTMs in biological samples using
stable isotopes [e.g. Isotope-coded affinity tags (ICAT),
stable isotope labeling with amino acids in cell culture
(SILAC), dimethyl labeling]

Absolute quantification of proteins using stable
isotopically labeled standards

Structural elucidation of small molecules
(e.g. metabolites) using liquid chromatography - accurate
mass measurements

MS analysis of chemically cross-linked peptides

Method development for proteomics (e.g. chemical
derivatization of PTMs)
Instrumentation:

Linear Ion Trap (LTQ) -Orbitrap Mass Spectrometer
coupled to nanoscale liquid chromatography

Two Linear Ion Trap (LTQ) Mass Spectrometers
coupled to nanoscale liquid chromatography
Personnel:
Dr. Ying Wai Lam, Director (left)
Tel: (802) 656-4709; E-mail: [email protected]
Dr. Bin Deng, Manager (middle)
Tel: (802) 656-5099; E-mail: [email protected]
Julia Fields, Technician (right)
Tel: (802) 656-1186; E-mail: [email protected]
F ACILITY
GUIDELINES
Sample Submission, and Data Management
T YPICAL W ORKFLOW
OF B OTTOM - UP P ROTEOMICS
Protein identification
Step 1: Initial consultation: Investigators should contact the
facility prior to sample submission to ensure a clear understanding
of sample requirements for processing through the facility. To
arrange a consultation, online consultation signups are available on
our website or you can contact Drs. Wai Lam / Bin Deng by phone
(802-656-4709, 802-656-5099) or email ([email protected],
[email protected]).
Step 2: Sample preparation: For gel samples, please provide an
image of the gel and indicate the bands or spots that are to be
analyzed. We need to be sure that your samples and sample
preparation methods are compatible with mass spectrometry and
that your sample has an adequate amount of the target protein(s) to
be measured by mass spectrometry. Usually, a visible silver or
Coomassie Blue-stained gel band/spot that corresponds to 20
femtomole - 1 picomole of protein is needed to identify the
protein.
PTM Analysis
Quantitative Proteomics
Using Stable Isotopes
For protein samples in solution, you must provide the expected
protein concentration and composition of the solution.
Samples submitted for phosphorylation analysis must be Coomassie
-stainable (in gel) or with a concentration higher than 5 pmole (in
solution).
Step 3: Fill out the sample submission form and answer
the questions listed: Each sample must be accompanied by a
completed form — please go to our website for online submission.
The information you provide will help us to ensure proper handling
and analysis of your samples.
Step 4: Analysis: Only trained personnel directly operate the
mass spectrometers in the facility. However, users are welcome to
learn and participate in data analysis.
Step 5: Results: Results will be sent via e-mail or provided as a
printed copy. Raw data will be provided upon request.
It is best to send your samples via overnight shipping if you are a
non-UVM user, otherwise please drop them off in person.
Please note that drop-off hours are Monday through Friday,
9:00 AM to 4:00 PM, and you are encouraged to give us a call
before dropping off. If you have any questions, please feel free to
contact us!
The higher the quality of your
samples , the quicker you will receive
your successful results!!!
Metabolite Identification