Gel Imaging and Analysis
Gel Imaging and Analysis
Phosphorimager Gel/Blot Analysis (1D and 2D)
For direct quantitation of radioactive bands in dried gels or blots, a BioRad Molecular Imager FX Pro Plus PhosphorImager is available, with a linear range of detected density of 5 orders of magnitude. Price is $2 per scan.
>> To schedule, contact Anne Stanley (x6087, astanley@psu.edu) or Suja Maddukuri (x6087, srm25@psu.edu).
Fluorimager Gel/Blot Analysis (1D and 2D)
The FX Pro Plus also has a 3-laser Fluorimager (488, 532, & 635 nm lasers) which enables band and spot detection from a large variety of fluors. After image acquisition, Quantity One 1D or PDQuest 2D analysis software can be used to perform calculations of band densities, background corrections, etc. The PDQuest software can also compare spot densities on different gel images to select protein spots whose levels change between different experimental conditions, and a spot cut list can be exported to our LEAP/BioMachines 2DiDx gel spot cutter. The imaging charge is $2 per scan, and the instrument is available on a sign-up basis to individual trained users.
>> To schedule, contact Anne Stanley (x6087, astanley@psu.edu) or Suja Maddukuri (x6087, srm25@psu.edu).
Gel/Film Densitometry Analysis (1D and 2D)
For quantitation of exposed film (radioactive or chemiluminescent detection), a BioRad GS800 Calibrated Densitometer is available to digitize film images. The densitometric image obtained can then be analyzed using Quantity One software to analyze 1D gels or slot/dot blot images, or PDQuest software to analyze two-dimensional gel images. The rate for Densitometry is $5/hr, and the instrument is available to trained users on a sign-up basis.
>> To schedule, contact Anne Stanley (x6087, astanley@psu.edu) or Suja Maddukuri (x6087, srm25@psu.edu).
DIGE Gel Analysis (2D)
For quantitation of differences in protein amount between different samples, equal amounts of each sample can be labeled with different fluors, usually a combination of Cy2, Cy3, and Cy5, then mixed together before 2D gel separation. Fluorescent imaging and quantitation of the resulting 2D gels can be done with our Typhoon Imager, and spot-cut lists can be exported for automated gel spot excision using our Ettan Spot Picker.
>> To schedule, contact Dr. Bill Freeman (x4037, wfreeman@psu.edu )