成体脳におけるセマフォリン3A発現を定量するELISAシステムの確立

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  • Construction of the ELISA assay to quantify Semaphorin 3A in the adult brain

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<p>Extracellular soluble signals that control several aspects of neuronal development are known to play a critical role in maintaining neuronal function and homeostasis in the mature nervous system. Abnormal expression and/or secretion of these molecules are therefore thought to be associated with the onset of various types of neurological disorders. It has been reported that the expression of Semaphorin 3A (Sema3A), a secreted type of repulsive axon guidance molecule, is impaired in several neurodegenerative disorders. However, due to the lack of a reliable Sema3A antibody, our knowledge about Sema3A expression in the adult brain is still limited. Here we report the identification of a pair of Sema3A monoclonal antibodies for the sandwich ELISA assay using the Autonomously Diversifying Library system. This ELISA assay recognized recombinant human Sema3A in the range of 0-100 pM, but not Sema3F, one of a subfamily of class III semaphorins. The specificity of this assay was also confirmed by using the embryonic brains from sema3A deficient mice as a negative control. Moreover, this assay could measure Sema3A concentration in TBS- and RIPA-soluble lysate obtained from the adult mice brains. These data suggested that our ELISA assay is a reliable tool for the validation of Sema3A as a biomarker in neurodegenerative disorders.</p>

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