The CCIC-Mass Spectrometry and
Proteomics Facility is undergoing a major expansion in partnership
with the College of Medicine to add high throughput proteomic
research capabilities to the Facility. The expansion is currently
adding an Ettan Spot Handling Workstation and Typhoon Variable Mode
Image Scanner worth $600K. The Ettan Spot Handling Workstation and
Typhoon Image Scanner is a fully integrated instrument for automatic
processing visualization and transfer of proteins from
polyacrylamide electrophoresis gels for direct high throughput mass
spectral analyses. The expansion and acquisition of the new
instrumentation will make The Ohio State University Mass
Spectrometry and Proteomic Facility a premier research Facility in
the Country.
The
Ohio State University Mass Spectrometry and Proteomics Facility has
considerable experience in identifying proteins from 1D and 2D gels
using the current state-of-the-art mass spectrometers for proteomic
analysis. The methods available to ID proteins in our facility is
quite sensitive and can be used to identify proteins in the 10-50 ng
range for bands detected by zinc, SYPRO ruby or Coomassie staining.
Enzymatic digestion of proteins, (in solution or in-gel) followed by
MS/MS sequencing of the resulting peptide mixture leads to sequence
tag.
Using this procedure, an unknown isolated protein can be enzymatically digested by a sequence specific protease, such as
trypsin. The resulting sequence tag is then BLAST searched for
protein ID. We have found this method to be sensitive and powerful,
producing numerous sequences from 200 fmole of material and enabling
us to identify proteins for which peptide mass mapping was
unsuccessful.
Our
Facility has recently installed a Dionex LC-Packings Capillary LC
system. The Capillary LC system equipped with 2-D LC-chromatography is a critical analytical tool for
your proposed research as it allows analysis and identification for
complex protein mixtures (MudPIT).
Due to recent guideline for protein identification published in
Molecular and Cellular Proteomics (2004, 3.6 page 531) we
are only identifying proteins using MS/MS sequencing. Protein ID by
MALDI will only be considered in special circumstances. As always
our staff will make every effort to provide you with the highest
quality results possible.
Please contact Dr.
Kari B. Green-Church (614.688.0521) for sample preparation
advice to ensure quality results are obtained.
Sample Preparation Guidelines for 2-D
Careful preparation
of your sample can be the difference between positive and negative
results. Consult a MS&P professional if you are unsure about your
sample preparation.
Click on the image for
a Power Point Presentation of Proteomics 101.
