Peptide Research Protocols: Endothelin (Methods in Molecular by Lorette C. Javois

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By Lorette C. Javois

A panel of multidisciplinary specialists describes intimately without problems reproducible the way to examine all facets of the endothelin procedure from its synthesis and metabolism, to its functionality in well-being and illness. Theses equipment use state of the art molecular options to quantify the expression of mRNA for either endothelin receptors and the endothelin changing enzymes. They express how peptides, precursors, receptors, and artificial enzymes might be localized and quantified in plasma, tradition supernatants, tissue homogenate, and tissue sections utilizing antibodies. numerous in vivo protocols illustrate the function of the endothelin peptides in fit human members and describe animal versions that may be used to foretell the healing strength of cardiovascular medicines that manage endothelin synthesis or functionality.

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This limitation must be considered in the design of experiments to ensure appropriate control to allow measurement of relative changes rather than absolute levels of immunoreactivity. Sandwich ELISAs are available from a number of commercial sources including the Biotrak™ endothelin-1 ELISA from Amersham Pharmacia Biotech (19). This kit, one of the first to be developed, is recommended because of excellent intra- and inter-assay coefficients of variation and good 24 Davenport and Kuc recovery following solid phase extraction (19).

Vasoactive intestinal contractor, the rodent (mouse and rat) equivalent of human ET-2, differs by only one amino acid. The assays described below would be expected to detect the mature peptides in most, if not all, mammalian species. , fish), the N-terminus is reported to vary from the human ET-1 sequence but the C-terminus is conserved; selecting antisera directed to the C-terminus should therefore ensure 100% crossreactivity. The C-terminus of big ET precursors can also vary between species. 5.

1994) Effects of phosphoramidon and pepstatin A on the secretion of endothelin-1 and big endothelin-1 in human umbilical vein endothelial cells: Measurement by two-site ELISAs. Clin. Sci. 87, 245–251. 20. Davenport, A. , Ashby, M. , et al. (1990) A sensitive radioimmunoassay measuring endothelin-like immunoreactivity in human plasma: comparison of levels in essential hypertension and normotensive controls. Clin. Sci. 78, 261–264. 21. , Haynes, W. , Webb, D. , and Davenport, A. P. (1995) Phosphoramidon inhibition of the in vivo conversion of big endothelin-1 to endothelin-1 in the human forearm.

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