| Single Gel / Three Images | |
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Each protein sample
is labeled with either Cy2, Cy3, or Cy5 cyanine dyes,
combined, and then resolved on a single two-dimensional gel. A digital image is generated
from each Cy dye. In this experimental design, the uncertainties of
feature matching are eliminated and manual image analysis is minimized.
The first sample (S1) may be a mixture of standard proteins (provided by the Proteomics Core), a pooled standard generated by combining all of the samples in a multi-gel study (to match the images from this gel with others in the study), or another protein sample for direct comparison to samples S2 and S3. Images are produced and analyzed using DeCyder Differential Analysis software (Amersham BioSciences). Click here to submit samples for 2D gel analysis |
| Multi-Gel Analysis / Pooled Internal Standard | |
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Multi-gel comparisons may be performed
using different combinations of sample sets. In the example a
control sample (C1)
is labeled with Cy 3, and samples S1,
through Sn are
labeled with Cy 5. A pool of all samples is labeled with Cy 2
and serves as a standard common to each gel. The pooled
standard, the control, and one of the test samples are combined and
run on each gel. Images are generated, and
compared within each two-dimensional gel (using DeCyder DIA image analysis software) and
between gels (using other image analysis software) to
quantitate relative differences among the samples. Inter-gel analysis is considerably more
time-consuming and inexact than single gel analysis. When possible,
single gel three image pilot analyses are recommended.
Click here to submit samples for 2D gel analysis
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Fees are for Washington University investigators. Non-WU investigators should contact Jim Malone at (314) 286-2452 for pricing.
To schedule an appointment to discuss an experimental design call (314)286-2452
