Bio-Rad EXQuest Spot Cutter - Manual (Page 13)

PDQuest User Guide
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Sample Experiments
Valuable information can be learned by exposing cells to a set of specific
experimental conditions and subsequently examining their biological response.
A preliminary in vitro experiment that is useful when beginning Raw 2-D gel work is
to radiolabel a cell’s proteins to steady-state with
35
S-methionine. Running gels of
these proteins will help to determine the commonly expressed proteins in the cell
under normal conditions.
Subsequently, other amino acids such as
3
H-leucine,
3
H-proline,
3
H-lysine, etc., can
be used to radiolabel the proteins to steady-state. This will label any proteins that do
not contain methionine and were, therefore, not detected in the first experiment, and
will provide preliminary information on the amino acid composition.
Other experiments complementary to Raw 2-D gels include: cell fractionation
procedures, and post-translational modifications (phosphorylation, methylation, etc.).
Data from these experiments can be added to a database, accumulating information
about these proteins.
3. Bio-Rad Listens
The staff at Bio-Rad are receptive to your suggestions. Many of the new features and
enhancements in this version of PDQuest are a direct result of conversations with our
customers. Please let us know what you would like to see in the next version of
PDQuest by faxing, calling, or e-mailing our Technical Services staff. You can also
use Solobug (installed with PDQuest) to make software feature requests.