University of Vermont Proteomics Core Facility:
Current and Future Capabilities & Services
|
Current Proteomics Facility Capabilities |
Dependence on User Protocol |
Amount of Submitted Sample |
Required Time* |
Protein identification by peptide mass fingerprint using MALDI-TOF-MS (limit: single protein) |
Moderate |
> 1 pmol |
1 day |
Protein in-solution digestion and identification by LC-MS/MS |
Low |
> 100 ng |
2.5 days |
Protein in-gel digestion and identification by LC-MS/MS |
Low |
> 1 µg |
3 days |
Identification of proteins in a mixture by LC-MS/MS (limit: < 200 proteins in a mixture) |
Moderate |
> 1 pmol |
2.5 days |
Identification of protein posttranslational modifications: phosphorylation, methylation . . . |
High |
> 5 pmol |
1 week |
Determination of protein complexes from client-supplied protein mixtures |
High |
> 1 pmol |
1 week |
Identification and quantification of specific peptides from proteins using selected ion monitoring |
High |
> 500 amol |
1 day |
Assistance with system biology methods |
High |
|
2 days |
*Assumes properly prepared samples in adequate amounts and in simple (i.e. LC/MS-friendly) sample matrices.
VGN PROTEOMICS FACILITY—PRICING
Below is the pricing schedule for protein samples (bands or spots or regions of gels) excised from polyacrylamide gels. However, should you find the prices prohibitive please contact VGN directly for a discussion of your needs (Vermont.GeneticsNetwork@uvm.edu).
| Subsequent gel band/spot: | |
Silver-Stained Gel Bands/Spots |
$25 per gel band/spot |
Coomassie-Stained Gel Bands/Spots |
$10 per gel band/spot |
Pricing for large projects (more than 20 gel bands/spots at a time) will be by quote. (contact: Bin.Deng@uvm.edu) |
|
Although gel-based samples are typically the most straight-forward and the cleanest to analyze, it is understood that some proteomic projects will not involve gels, and pricing for such projects will be by quote (contact: Bin.Deng@uvm.edu).
In general, coomassie-stained gel bands/spots contain protein levels that will greatly enhance proteomic success. When possible, we strongly encourage the submission of coomassie-stained gel bands/spots. In addition, some silver-staining procedures are much less compatible with mass spectrometry than others. Commercial kits are available that are mass spectrometry compatible and include G Biosciences FOCUS-Fast Silver or FAST silver Staining Kits and Invitrogen's SilverQuest Silver Staining Kit: Please see the VGN website below for an example of a generic mass spectrometry-friendly silver staining protocol:
Silver Staining of Polyacrylamide Gels
To set up a required pre-submission consultation with the proteomics staff, please follow the links below:
Future Proteomics Facility Capabilities |
Identification of proteins and peptides by static nano-ESI |
2D-LC-MS/MS for complex protein mixtures coming from cell lysates, tissue, and blood samples |
Protein molecular weight determination by MALDI-TOF-MS and ESI-MS |
Off-line separation and fraction collection from HPLC: peptide and protein purification and SCX-HPLC |
Phospho-peptide specific enrichment and identification from complex mixtures |
Structure analysis by MSn |
Identification of lower abundance plasma proteins after depletion of high abundance proteins |
Quantification of peptides and proteins using stable isotope tags |
Identification of glyco-proteins |
