111 Glaxo Building
101 Mason Farm Road
Campus Box 7028
Chapel Hill, NC 27599-7028
The proteomics facility offers 2 levels of protein identification services. All protein identification services include searches of publicly available databases and a results report. Database searches of client-provided sequences or specific modifications are available for an additional charge.
Identification of gel-separated proteins
Protein bands or spots separated by 1-D or 2-D SDS-PAGE are excised and digested in-gel (typically with trypsin, other enzymes available if needed upon consultation) and the resulting peptides are analyzed by ABI 4800 MALDI TOF/TOF MS. Charges include a setup fee for in-gel digestion and a fee per individual protein band or spot analyzed.
Identification of proteins or peptide mixtures
The facility offers larger scale analysis of proteins in solution derived from co-immunoprecipitation or similar experiments. Proteins of interest are digested in-solution and the resulting peptides are analyzed by LC-MALDI-TOF/TOF. Services include database searching and protein identification reports and simple C4 or C18 sample desalting.
Analysis of Post-translational Modifications (PTMs)
The facility offers protein phosphorylation analysis of purified proteins or proteins derived from 1-D or 2-D SDS-PAGE. For proteins from SDS-PAGE an in-gel digestion setup fee applies. All other proteins are digested in-solution and peptides analyzed by ThermoScientific LTQ OrbiTrap LC-MS. Services include database search and reporting as well as simple C4 or C18 sample desalting.
Mass Determination of Peptides or Proteins
The proteomics facility offers mass analysis of purified peptides and proteins by ABI 4800 MALDI TOF MS. For proteins, low mass accuracy and resolution can be obtained using our MALDI-TOF/TOF in linear ion mode and high mass accuracy, analysis by ESI-LTQ Orbitrap is required.
Training on ABI 4800 MALDI TOF/TOF
Training will include instructions on the preparation and analysis of proteins and peptides by ABI 4800 MALDI TOF/TOF mass spectrometry. The basic principles of MALDI analysis (both MS and MS/MS), instrument calibration and usage, method set-up, sample analysis and data analysis will be included. Once trained, the user will be able to calibrate the instrument, set up MS and MS/MS “jobs” and perform database searches (GPS Explorer/ Protein Pilot) to obtain the results of these analyses. Time: approximately 4 hrs total (split over more than one day).
Training on Tempo-LC with spotting onto MALDI plates
This training is for advanced users only and prior MALDI TOF/TOF training is required. Training will include sample preparation and instruction in instrument operations. Trained individuals will know how to utilize software, inject samples, and maintenance and operate LC. Time: 8 hrs total (split over multiple visits).
Mascot/SEQUEST* Software Training
This will include additional training in the Mascot database and Sequest search algorithms. Time: approximately 1 hr.
* Mascot is a registered trademark of Matrix Science Limited. SEQUEST is a registered trademark of the University of Washington.
(Subject to change)
Fee Schedule Services UNC Non-Profit For Profit In-Gel band digestion setup fee (per gel) $100 $150 $200
Protein ID by MALDI-TOF/TOF (per band/spot)
50 75 100 LC spotting onto MALDI plate with MALDI-TOF/TOF Analysis 400 600 800 Additional Sample Analysis (per Hour) 50 75 100
Peptide Mass Measurement (per sample)
20 30 40 Peptide Mass Measurement (per plate - 30 spots maximum) 100 150 200 PTM Analysis by Orbitrap (per sample) 500 750 1000 Sequence Analysis (de novo) 100 150 200 Solid phase purification of peptides (up to for samples) 200 300 400 Solid phase purification of peptides- training 100 N/A N/A Solid Phase purification of peptides - Supplies 50 N/A N/A iTRAQ labeling 450 675 900 Analysis of Complex Mixture 400 600 800
In depth analysis of Protein Mixture (Peptide fractionation - 2D LC or equiv)
3000 4500 6000 Peptide Purification (zip tip) 50 75 100 4800 training 300 N/A N/A Tempo LC Training * N/A N/A Orbitrap Training * N/A N/A Mascot Training 50 N/A N/A Instrument Usage - 4800 (hand spotting) (First Hour) 50 75 N/A 4800 Instrument Usage - (Automatic Spotting) (Per Hr. of Analysis) (overnight) 30 N/A N/A Instrument Usage - Orbitrap (per 90 min. Gradient) 250 N/A N/A Low Resolution Protein Mass Measurement § (First Sample) 100 150 200 Low Resolution Protein Mass Measurement § (Additional Samples) 30 45 60 High Resolution Protein Mass measurement (First Sample) 150 225 300 High Resolution Protein Mass measurement (Additional Samples) 50 75 100
N/A – not available
* - Currently, not available. In the future we would like to make these available.
§ - This is for protein mass measurment. If your compound is not a normal protein, please provide a reference for the MALDI-based method used by others. We have poor success without this information because it is a matter of trial and error and we cannot devote extensive time for this type of analysis.