AfCS > AfCS laboratories > Antibody Laboratory
ALLIANCE FOR CELLULAR SIGNALING
ANTIBODY LABORATORY
University of Texas Southwestern Medical Center
Dallas, TX
Overview
The primary goals of the Alliance for Cellular
Signaling (AfCS) Antibody Laboratory are to quantify ligand-induced changes
in the secretion of cytokines and in site-specific phosphorylation of
signaling proteins of interest. These projects constitute part of the AfCS
ligand screen—an effort to apply several assays to define the pattern of
cellular response to ligands and to identify nonadditive interactions between
combinations of ligands. Groups of (4 to 6) phosphospecific antibodies are
employed to monitor simultaneously the phosphorylation of specific sites in
several proteins by multiplex Western blot technique. These proteins are
chosen to represent several signaling pathways and ones in each group are
resolved on a single lane of a gel. To reduce reliance on labor intensive
Western blotting, the Antibody Laboratory is collaborating with Bio-Rad and
Cell Signaling Technology to expand the number of assays available to
monitor site specific protein phosphorylation by liquid suspension array
(Bio-Plex). This technology is state-of-the-art for measurement of cytokines
and is currently employed by the Antibody Laboratory.
Responsibility for acquisition and
characterization of antibodies needed for ongoing AfCS projects falls to the
Antibody Laboratory. Antibodies are procured from commercial sources or
produced on site or by commercial contract. The lab has developed
collaborations with antibody-producing companies that will both help to
contain AfCS costs and facilitate introduction of new reagents to the market
for the entire research community. The Antibody Laboratory has tested
numerous commercially available antibodies and has tabulated results in
database form. The database is a updated periodically and is a resource for
the research community. It is
available online in the Data Center of the
Nature's Signaling Gateway website.
Collaborators
The Antibody Laboratory is collaborating with Cell Signaling Technology (use
and development of antibodies)
www.cellsignal.com and
Bio-Rad Laboratories, Inc. (development of Luminex technology for monitoring
phosphoproteins) www.Bio-Rad.com.
Staff
Alliance for Cellular Signaling
Antibody Laboratory
University of Texas Southwestern Medical Center
5323 Harry Hines Blvd., J5-130
Dallas, Texas 75390-9196
Phone: 214-648-2271
Fax: 214-648-5006
| Director | Susanne Mumby, Ph.D. |
| Lead Scientist | Heping Han, M.D., Ph.D. |
| Research Scientist | Ruth Levitz, Ph.D. |
| Research Assistant | Eduardo Arteaga |
| Research Assistant | Nicholas Wong |
| Research Assistant | Ila Oxendine |
| Research Assistant | Frank Amador |
| Research Assistant | Rod Ceja |
| Research Assistant | Blythe King |
| Research Assistant | Justin Hill |
| Research Assistant | Ari Malka |
| Lab Administrator | Kimberly Edwards |
| Data Manager | Lonnie Sorrells |
| Lab Aid | Philip Ramirez |
Slide Presentations from May 23-26, 2004, AfCS Annual Meeting
Goals for Year 5 (9/04-8/05)
- Complete the RAW cell ligand screen for phosphoproteins
- Monitor site-specific protein phosphorylation by Western blotting
for experiments that are suggested from analysis of the double ligand
screen, including perturbation studies
- Continue to pursue development of suspension array (Luminex)
assays for phosphoproteins
- Assay cytokines in experiments that include exposure of
cells to triple ligands or inhibitors of transcription and translation
- Provide cytokine data for perturbation studies, including knockdown experiments
- Continue to maintain the AfCS Antibody Database and provide
updates for public web display
