Titlebook: Membrane Transporters in Drug Discovery and Development; Methods and Protocol Qing Yan Book 2010 Humana Press 2010 In silico.PCR.bioinforma

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Measurement of Intracellular pH,pH is of interest in studying many processes, including pH regulatory transport proteins. Key approaches that have been used to determine intracellular pH include pH-sensitive microelectrodes, nuclear magnetic resonance, and pH-sensitive fluorescent proteins. Here we review these approaches while pr

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,Measurement of Plasma Membrane Calcium–Calmodulin-Dependent ATPase (PMCA) Activity,racellular compartment. Although in excitable cells such as skeletal and cardiac muscle cells PMCA has been shown to play only a minor role in regulating global intracellular calcium concentration, increasing evidence points to an important role for PMCA in signal transduction, in particular in the

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Methods to Evaluate Transporter Activity in Cancer, of delivering a drug to the target tissues, avoiding distribution to other tissues, and improving oral bioavailability. This chapter focuses on methodology to analyze the activity of transporters that are involved in drug transport.

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Site-Directed Mutagenesis in the Study of Membrane Transporters,res for many membrane transporters other molecular tools need to be employed. In vitro site-directed mutagenesis is one method that has the capacity to provide both structural information and identification of the role of individual residues and/or regions of a protein that are involved in function.

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, Oocytes,the analysis of membrane transporters in this system, including preparation of oocytes, assays of transport activity, protocols for immunostaining and fluorescence microscopy, and other assays to study surface expression.

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Measurement of Intracellular pH,r pH include pH-sensitive microelectrodes, nuclear magnetic resonance, and pH-sensitive fluorescent proteins. Here we review these approaches while providing details on the use of pH-sensitive fluorescent dyes to measure cytosolic pH.