Titlebook: Neurotrophin Protocols; Robert A. Rush Book 2001 Springer Science+Business Media New York 2001

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Localization of Neurotrophin Proteins Within the Central Nervous System by Immunohistochemistryeaching an understanding of endogenous neurotrophin function. One technique for addressing many of these issues is immunohistochemistry. The strength of the immunohistochemical approach lies with its ability to precisely define the distribution of a given protein within a tissue sample obtained unde

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Neurotrophin Immunohistochemistry in Peripheral Tissuesal intervention. The movement of the neurotrophins has been studied extensively by the use of nerve ligations, which allows determination not only of the retrograde and/or anterograde movement of the protein, but also can be useful in demonstrating the absence of a neurotrophin in a particular class

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Quantification of mRNA Levels Using Ribonuclease Protection Assaym of a sequence that encodes a portion of neurotrophin mRNA. This DNA fragment can either be part of a cDNA clone or a genomic subfragment, but it should be selected so that the plasmid construct can be digested with a specific restriction enzyme prior to transcription, to yield a labeled RNA specie

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Quantification of Neurotrophin mRNA by RT-PCRrsatility, where total RNA is first reverse transcribed and then amplified by PCR reaction (.,.). The technique can be used to detect the presence or absence of gene transcripts, to measure expression levels semiquantitatively for comparison of mRNA levels in different tissues, or to determine the a

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,,oles of neurotrophins are often closely related to their gene expression during development. Detection of their mRNA expression in developing and in experimental animals has accumulated considerable data revealing many biological functions of these molecules in the normal development and in patholog

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Construction of Cells Expressing Neurotrophinsarge precursor protein within which mature, bioactive NGF occupies the C-terminal half, and the N-terminal half contains a signal peptide (18 amino acid) preceding a large pro region. The pro sequence in combination with the signal (pre) sequence is necessary for the secretion of correctly processed

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