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Mass Spectrometry Facility

Mass Spectrometry Facility

Director:  Lauren E. Ball, Ph.D.; 843-792-4513, ballle@musc.edu

Lab Manager:  Jennifer Bethard, M.S.; 843-792-8637, bethard@musc.edu

Staff:  Susana Comte-Walters and Kim Norris-Caneda

Location: 305 Children’s Research Institute

 

Description:  This Core is housed within the MUSC Mass Spectrometry Facility which provides expertise, services, education, and training to enhance biomedical research endeavors through mass spectrometry-based proteomics. Currently there are over 50 investigators which utilize the facility for protein identification and characterization. Protein analysis includes in-gel or in-solution protease digestion, chromatographic separation and tandem mass spectrometric analysis of the resulting peptides, and interpretation of MS/MS data using Sequest , Mascot,  Protein Pilot, MaxQuant, and other search algorithms. The facility also assists in the development of customized applications for the isolation, detection and characterization of posttranslationally modified peptides (e.g. phosphorylation, glycosylation, oxidation, glutathionylation, and O-GlcNAc modification). With the recent acquisition of the Orbitrap Elite Mass Spectrometer we are expanding our services to couple quantitative approaches (SILAC, iTRAQ®, ICAT®, TMT®) to modification-specific experiments (eg., phosphoproteomics, redox proteomics). We are developing methodology to analyze alterations in posttranslational regulation that impact signal transduction, epigenetic modulation, and the response to therapeutics with the goal of enabling investigators to discover molecular mechanisms underlying disease progression and therapeutic responses that may not be revealed through genomic studies.

Services: MALDI-TOF MS, LC-MS, and LC-MS/MS tandem mass spectrometry analyses are offered for protein analysis.  Protein identification services include in-gel or in-solution protease digestion, chromatographic separation and tandem mass spectrometric analysis of the resulting peptides, and interpretation of MS/MS data using Sequest® or Mascot® software. The facility will also assist in the development of customized applications for the isolation, detection and characterization of posttranslationally modified peptides (e.g. phosphorylation, glycosylation, oxidation, glutathionylation, and O-GlcNAc modification).  Sites of modification are verified by manual inspection of the data.  Please consult facility staff for feasibility and pricing of quantitative proteomic experiments (iTRAQ or SILAC), the implementation of specialized approaches with quantitative proteomics (e.g. phosphoproteomics, O-GlcNAc proteomics), and MALDI-imaging mass spectrometry for tissue imaging experiments.

Services ProvidedCost
  
Protein Identification 
MALDI TOF/TOF (gel spot/band)$100/sample (Digestion included)
LC-MS/MS (gel spot/band) fast- gradient$150/sample (Digestion included)
LC-MS/MS moderately complex mixture-medium gradient$175/sample (Digestion included)
LC-MS/MS complex mixture or PTM ananlysis - long gradient$200/sample (Digestion included)
2D LC-MS/MS (5 fractions) ($125/salt step/sample)$625 (Digestion Included)
2D LC-MS/MS (10 fractions) ($125/salt step/sample)$1250 (Digestion Included)
MW Determination by MALDI-TOF MS$25 (1-3 samples) $50 (4-6)
  
Single Protein Characterization 
Identification of Post-Translational Modifications$200/LC-MS/MS analysis
Quantitation of Relative Changes in Post-Translational Modifications

Includes comparison of 2 treatment conditions with 3 replicates each (label free). Trypsin digestin, 6 LC-MS/MS analyses.
$1,200
Phosphorylation enrichment and analysis$240/sample
De Novo Sequencing/manual interpretation of analysis$50/hr
  
Quantitative Proteomics (Orbitrap Elite Mass Spectrometer) 
Differential Protein Expression 

SILAC

2-3 Treatment conditions, gel fractionation, protein digestion, 12 LC-S/MS analyses for each biological replicate
$4,800/2 biological replicates

TMT6 plex, TMT10 plex, iTRAQ 4 plex (Isobaric tagging)

Includes reagents, digestion, labeling, fractionation, and 12 LC-MS/MS analyses
$4,350/experiment

Label Free Proteomics (MaxQuant LFQ-Quantitation based on chromatographic peak areas)

Includes 2 treatment conditions, each with three technical replicates.

$3,200/experiment
Regulated Sites of Post-translational Modifications: 

Phospho SILAC

Includes digestion, SCX or bpH-RP fractionation, TiO2 enrichment, and LC-MS/MS analysis of 12 fractions.

$5,200/biological replicate

Acetyl SILAC

Includes digestion, fractionation (10 fractions), acetyl peptide enrichment, and 10 LC-MS/MS analyses

$6,550/biological replicate
*cost includes antibody
*Prices current as of September 2015

Consultation: Please contact any of the listed personnel for questions and consultations regarding experimental design and proteomic analysis options.

Instrumentation:

Mass spectrometers and associated proteomic applications available include:

• Bruker Solarix 7T Dual Source MALDI/ESI FT-ICR MS (CID and ECD Fragmentation)
MALDI Tissue Imaging, Top-Down Protein Characterization

• Thermo Orbitrap Elite with VelosPro Ion Trap MS (CID, HCD, ETD Fragmentation)
LC-MS/MS for identification, characterization of modifications, quantitation of differential protein expression or posttranslational modification using SILAC, iTRAQ®, TMT® , or label free approaches. Top-Down Protein Characterization.

• Thermo LTQ XL Linear Ion Trap MS (CID, PQD, fragmentation)
LC-MS/MS for protein identification and characterization of fragile modifications.

• Applied Biosystems 4800 MALDI-TOF-TOF Proteomics Analyzer
LC-MALDI-MS/MS for protein identification and quantitation of differentially expressed protein class="bodytext"using iTRAQ®, or TMT®  reagents.

• Bruker Autoflex III MALDI-TOF-TOF MS
MALDI Tissue Imaging

• Associated HPLC systems (5 LC Packings nano-LC systems and 2 Dionex Probot MALDI Spotters for LC-MALDI)

 
 
 

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