Resources

Our current resources include a variety of mass spectrometers interfaced to nanoHPLC, a SPR Biosensor (Biacore), 1D and 2D SDS-PAGE equipment, and an AKTA protein purification instrument.

 

LTQ-Orbitrap (2010) (Financial support provided by NIH/NCRR 1 S10 RR028868-01)

 

G Tsaprailis and Orbitrap LTQ

Dual-Pressure Linear Ion Trap with Orbitrap mass analyzer
Resolving power of >100,000
Mass accuracy better than 1 ppm
Multiple fragmentation techniques: CID, HCD, and ETD
ETD shown to maintain PTMs intact for facile mapping
Parallel MS and MSn analysis
Advion Nanomate ESI source for static ionization (96 well plate format sampling), nanoLC flow coupling ionization, and fraction collection
Sensitivity in low fmol range
Top-down sequencing of small to medium sized proteins
Proxeon nano HPLC with cooled autosampler (250 - 750 nanoL/min flow rates using 75 um columns)

 

LTQ (2006) (Financial support provided by Ventana Medical, Tucson AZ)

2D quadrupole ion trap mass spectrometer
ESI (nanospray)
MS and MS/MS
Nano LC-MS/MS system
Flow rates 250 nanoLit/min
Michrom Paradigm MS4 quternary pump HPLC
TSP 3000 autosampler
Fused silica (100 um ID x 365 um OD) reversed phase (RP) C18 columns (tip 3-5 um); self packed to 7 cm
Protein identification applications
Sensitivity 30-50 fmol standard digest (bovine serum albumin)

LTQ (2006) (Financial Support provided by NIH U54-A1065359)
2D quadrupole ion trap mass spectrometer
ESI (nanospray)
MS and MS/MS
Nano LC-MS/MS system
Flow rates 250 nanoLit/min
Surveyor quaternary pump HPLC
Surveyor autosampler
Fused silica (100 um ID x 365 um OD) reversed phase (RP) C18 columns (tip 3-5 um); self packed to 7 cm
Protein identification applications (A)
Multidimensional Protein Identification Technology (MudPIT) or LC-LC-MS/MS(B)
Fused silica (250 um ID x 365 um OD) strong cation exchange (SCX) Whatman Partisphere self packed to 5 cm
Whole proteome analyses
Solution protein mix
Gel separated protein mix (whole lane)
Switch between (A) and (B) design as dictated by necessity
Sensitivity 30-50 fmol standard digest (bovine serum albumin)

ABI 4000 QTRAP (2006) (Financial support provided by NIH 1S10RR022384-01)

Hybrid triple quadrupole linear ion trap mass spectrometer
ESI and nanospray
MS and MS/MS
Single ion monitoring (SIM), multiple reaction monitoring (MRM), neutral loss scan
Nano LC-MS/MS system
Flow rates 250 nanoLit/min
LC Packings Ultimate dual pump HPLC
Dionex FAMOS autosampler
Fused silica (100 um ID x 365 um OD) reversed phase (RP) C18 columns (tip 3-5 um); self packed to 7 cm
Small molecule analyses
Protein characterizarion
Targeted MS/MS and MRM workflow

Waters Q-TOF Premier (2008) (Financial support provided by NIEHS ES06694, the University of Arizona and the Arizona Cancer Center)

Qtof

ESI and nanospray
MS and MS/MS
Nano LC-MS/MS system
Waters nanoACQUITY
Label free quantitation in liquid phase
Ion Accounting software

 

 

Biacore T100 Biosensor (Funds were generously donated by “Our Friends in Prescott”: Jim and Linda Lee, Swayze and Laura McCraine, and Ron and Kathy James, to the Sarver Heart Center and to Dr. Carol Gregorio)

Biacore T100

Interaction information (Protein:Protein, Protein:Nucleic acid, Protein:Small molecule, Nucleic acid:Small molecule)
• Quantitative kinetic analysis (rate constants)
• Quantitative determination of affinity constants
• Concentration determination
• Determination of binding specificity (yes/no)
• Thermodynamic measurements

ABI Voyager DE-STR (2004) (Financial support provided by the University of Arizona and the College of Pharmacy)

MALDI ionization
MS
Linear mode
Refletron mode
Limited post-source decay (PSD)
Protein mass measurements
1 pmol/uL detection limit
Resolution 100-500
Peptide mass mapping
50 fmol/ul
Resolution 20000
MASCOT search engine

AKTA FPLC (2003) (Financial support provided by the BIO5 Institute)

 

 

 

 

 

 

 

Michrom HPLC (2007) (Financial support provided by Ventana Medical Systems, Tucson, AZ)

 

 

 

 

 

 

 

BioRad 1D and 2D SDS-PAGE equipment (2000-2007)

1D SDS-PAGE on 7 cm mini gels and 11 cm Criterion gels
Coomassie Blue or Silver stains
2D SDS-PAGE on 7 cm mini gels and 11 cm Criterion gels
variety of IPG strips for IEF in the 1st dimension (pH 3-10, 4-7, etc)

 

 

 

 

 

 

 

 

 

 

 

Sutter P200 Capillary puller (2000)

Fabrication of pulled fused silica columns (tips to 5 um opening) for nano C18 or SCX columns

 

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