Significant progress has been made toward specific aim 3. Currently, single adherent cells are lysed in less than 33 msec by a laser-induced shock wave. The contents of single cells are immediately loaded into a capillary, separated by electrophoresis within the capillary, and detected by laser-induced fluorescence. This was demonstrated and published in 1998 in Analytical Chemistry, vol 70, number 21, pages 4570-4577. Preliminary results show that kinase-substrate peptides can be measured from single mammalian cells. Further, cellular kinase activity can be measured as changes in the degree of substrate phosphorylation. Experiments are in progress to define parameters for and demonstrate physiologically relevant measurements.
Specific aims 1 and 2 are yet to be addressed substantively, but will be accelerated by the completion of the current studies on adherent cells.
Specific aim 4 will become possible immediately thereafter.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
3P41RR001192-20S1
Application #
6220404
Study Section
Project Start
1999-04-01
Project End
2000-03-31
Budget Start
1998-10-01
Budget End
1999-09-30
Support Year
20
Fiscal Year
1999
Total Cost
Indirect Cost
Name
University of California Irvine
Department
Type
DUNS #
161202122
City
Irvine
State
CA
Country
United States
Zip Code
92697
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