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Polyclonal antibody is produced by immunizing animals with a synthetic peptide surrounding Gln1875 of human Plexin A1 protein and purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly185 of human Plexin A2 protein, or Val440 of human Plexin A3 protein, or with a GST-fusion protein corresponding to residues from the human sequence of Plexin A4 protein.
Class 3 secreted semaphorin (Sema3A) is a chemorepellent that acts upon a wide variety of axons. As such, it induces a dramatic redistribution and depolymerization of actin filaments that results in growth cone collapse. Plexins are single membrane-spanning signaling proteins encompassing Plexin A1, A2, A3, and A4. Plexins form a complex with neuropilin-1 and -2 and the cell adhesion protein L1 to form a functional semaphorin receptor (1,2). The GTPase Rnd1 binds to the cytoplasmic domain of Plexin A1 to trigger cytoskeletal collapse. In contrast, the GTPase RhoD blocks Rnd1-mediated Plexin A1 activation and repulsion of sympathetic axons by Sema3A (3). Sema6A is a ligand for Plexin A2. Both Sema6A and Plexin A2 knock-out mice have a granule cell migration defect, where cells remain in the molecular layer. Furthermore, Plexin A2 also controls nucleus-centrosome coupling that modulates cell migration (4). Both Plexin A3 and A4 mediate the responses to class 3 semaphorins in sensory and sympathetic neurons. In particular, Plexin A4 is responsible for signaling of Sema 3A via neuropilin-1, while Plexin A3 is responsible for signaling of Sema 3F via neuropilin-2 (5).
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