The Production of Monoclonal Antibodies to VACM-1/Cullin 5
Dr. Vicki Isola
Vasopressin-activated calcium mobilizing receptor 1 (VACM-1/Cullin 5) is a 780-amino acid membrane protein that inhibits cellular proliferation and has been suggested to be a tumor suppressor. We propose to produce monoclonal antibodies to VACM-1/cul5. We will immunize Balb/c mice with purified VACM-1 and a KLH-conjugated peptide corresponding to the amino-terminal sequence of the protein. Spleen cells from mice expressing antibody titers over 1:30,000 will be fused with mouse myeloma cells, and hybrids will be screened for monoclonal antibody secretion using an ELISA assay. To date we have purified VACM-1 protein, prepared the conjugated peptide, and optimized the ELISA assay. Production of monoclonal antibody against VACM-1/cul5 will provide us with a novel tool to better characterize its expression in cancer cell lines and in tumor samples.
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