Anti-Phospho-ECT2 T790 Antibody

This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to an internal region of human ECT2 protein.

Expressed in lung epithelial cells (at protein level) . Expressed in squamous cell carcinoma, primary non-small cell lung cancer tumors and lung adenocarcinoma.

This affinity-purified antibody is directed against the phosphorylated form of human ECT2 protein at the pT790 residue. The product was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was first purified against the phosphorylated form of the immunizing peptide. The resultant affinity purified antibody was then cross-adsorbed against the non-phosphorylated form of the immunizing peptide. Reactivity occurs against human ECT2 protein at the pT790 residue. By ELISA the antibody is specific for the phosphorylated form of the protein. Reactivity with non-phosphorylated human ECT2 is minimal by ELISA, but western blot shows specific staining of interphase cell lysates, which are predicted to contain non-phosphorylated ECT2. Additional experiments are underway to clarify antibody specificity. A BLAST analysis was used to suggest cross-reactivity with ECT2 protein from human, mouse, rat, zebrafish, chimpanzee, chicken, and dog based on 100% homology with the immunizing sequence. Reactivity against homologues from other sources is not known.

Guanine nucleotide exchange factor (GEF) that catalyzes the exchange of GDP for GTP. Promotes guanine nucleotide exchange on the Rho family members of small GTPases, like RHOA, RHOC, RAC1 and CDC42. Required for signal transduction pathways involved in the regulation of cytokinesis. Component of the centralspindlin complex that serves as a microtubule-dependent and Rho-mediated signaling required for the myosin contractile ring formation during the cell cycle cytokinesis. Regulates the translocation of RHOA from the central spindle to the equatorial region. Plays a role in the control of mitotic spindle assembly; regulates the activation of CDC42 in metaphase for the process of spindle fibers attachment to kinetochores before chromosome congression. Involved in the regulation of epithelial cell polarity; participates in the formation of epithelial tight junctions in a polarity complex PARD3-PARD6-protein kinase PRKCQ-dependent manner. Plays a role in the regulation of neurite outgrowth. Inhibits phenobarbital (PB) - induced NR1I3 nuclear translocation. Stimulates the activity of RAC1 through its association with the oncogenic PARD6A-PRKCI complex in cancer cells, thereby acting to coordinately drive tumor cell proliferation and invasion. Also stimulates genotoxic stress-induced RHOB activity in breast cancer cells leading to their cell death.

Store vial at -20°C prior to opening. Aliquot contents and freeze at -20°C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4°C as an undiluted liquid. Dilute only prior to immediate use. Expiration date is one (1) year from date of opening.

This affinity-purified antibody has been tested for use in ELISA, western blot and immunoprecipitation. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 101 kDa in size by western blotting in the appropriate cell lysate or extract. Less than 2% reactivity is observed against the non-phosphorylated form of the immunizing peptide. This antibody is believed to be phospho specific for ECT2 phosphorylated at the pT790 residue. Experiments to generate more conclusive evidence are currently under preparation.

Nucleus. Cytoplasm. Cytoplasm, cytoskeleton, spindle. Cleavage furrow. Midbody. Cell junction. Cell junction, tight junction. Sequestered within the nucleus during interphase. Dispersed throughout the cytoplasm upon breakdown of the nuclear envelope during mitosis. Colocalizes with the centralspindlin complex to the mitotic spindles during anaphase/metaphase, the cleavage furrow during telophase and at the midbody at the end of cytokinesis. Colocalized with RhoA at the midbody. Its subcellular localization to tight junction is increased by calcium. Localized predominantly in the cytoplasm of numerous carcinoma cells.