Instrumentation

 

Mass spectrometers: New: Thermo QExactive Plus

LTQ-Orbitrap and QTRAP

Electrophoresis: 1D and 2D SDS-PAGE

Chromatography: HPLC, FPLC

 

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

George Tsaprailis and LTQ Orbitrap

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)

 

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


Q Trap and Yelena Feinstein

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 nL/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 workflows

  

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

FPLC and Cynthia David

Simultaneous variable UV-VIS wavelength, pH, and conductivity measurements
Designed for method scouting, development, and optimization
Two pump system with four buffer lines
Fraction collector has platforms for 96-well plates to 10 ml tube collection
 

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

HPLC and Cynthia David

Michrom Paradigm MS4 quaternary pump
Microbore plumbed
UV-VIS wavelength detector
Fraction collector

  

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

PAGE gels and Andrea Hunt

IEF and Michael Galligan

 

1D SDS-PAGE on 7 cm mini gels and 11 cm Criterion gels
Coomassie blue, silver, SYPRO Ruby, and ProQ 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)

capillary puller and Yelena Feinstein

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