Equipment available for protein analysis

The Biomolecular Research Facility has equipment which is either not commonly found in individual laboratories or is needed only occasionally by some laboratories. Some equipment is operated by facility personnel, and some is used by individual researchers. Much of this equipment has been mentioned elsewhere on this web site.

Fluorescent plate reader

Equipment is Gemini EM Spectramax. Reads microtiter plates with 96, 24, 12, 6 wells. 

  • dual monochromators allowing any fluorophore to be read within the  range of 250 to 850 nm 
  • fluorescent spectra capability
  • top or bottom reading of plates for adherent cells
  • well scanning
  • kinetic measurements
  • heating of plates
  • shaking of plate
  • SoftMax Pro 4.7 software for data analysis.
  • claimed sensitivity of 3 fmol/well of FITC with top reading 

Can perform luminescence readings, with less sensitivity but possibly wider range than a dedicated photon counting luminometer. Does not perform absorbance readings.
Operated by researcher. Fee for use. Contact person John Shannon  ph. 3-9399

High performance liquid chromatography.

Normally operated by facility personnel, fee for service.
Machines are: ABI 140 for 1 and 2 mm diameter columns, Beckman System Gold for 4.6, 10, 22 mm diameter columns. Detection by variable wavelength UV detector. Data collection by System Gold software.
Most often used for separation of peptides using reverse phase columns.
Contact people: John Shannon or Linda Beggerly  4-2869

Medium pressure chromatography using FPLC

Normally operated by facility personnel, fee for service.
The instrument is AKTA FPLC, detection at 280 nm (or 260), with a Frac-100 fraction collector. Data collected by UNICORN data system.
Columns used are gel filtration, ion exchange, for separation of proteins.
Contact person: John Shannon  ph. 3-9399

Circular dichroism, AVIV

Operated by researcher.
Contact person:John Shannon  ph. 3-9399

Dynamic Light Scattering.

Operated by researcher.
contact person: John Shannon  ph. 3-9399

Gel scanners

Bio-Rad GS800 for absorbance readings (visible wavelengths). Scanning area 290 mm x 400 mm. Absorbance or transmittance reading for gels and blots. Three wavelenght filters for different stains. Maxiumum spatial resolution 36.3 µm.

Bio-Rad FX fluorescent scanner, scan area about 290 mm x 400 mm. Three lasers and five filters to cover common fluorescent stains. Maximum spatial resolution 50 µm.

Both scanners produce files which can be converted to 8 or 16 bit tiff files. Bio-Rad Quantity One or PDQuest software can do some quantitation and comparison of images.

Spot cutter 

Both scanners can be used with a Bio-Rad spot cutter which illuminates gel (maximum area 220 mm x 200 mm) with either a UV light or visible light (no wavelength selection available).  A tip of 1 or 1.5 mm diameter cuts spots  (bands can be cut by making multiple spots) and deposits them into a 96 wel plate.

All these instruments are operated by facility personnel. Contact person John Shannon  ph. 3-9399