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Neuropeptide Protocols: Methods in Molecular Biology, Vol. 73

  •  ISBN-10 0-89603-399-6

  •  ISBN-13 978-0-89603-399-3

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publication info   G.B. Irvine and C.H. Williams, Humana Press, 1996, 378 pp., hard cover
mfr. no.   Humana

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Master protocols for analyzing neuropeptide structure and function. Techniques presented for extracting and purifying neuropeptides from a variety of biological sources for their sequence determination by Edman degradation and PCR. Also contains chapters on molecular modeling, radioisotopic labeling, production of antisera and the assay and inhibition of neuropeptidases.

Table of Contents

Preparation of Neuropeptide-Containing Fractions from Biological Materials, Purification of Extracted Peptides for Structural Analysis, Amino Acid Sequencing of Neuropeptides, Neuropeptide Gene Identification Using the Polymerase Chain Reaction. Solid-Phase Synthesis of Neuropeptides by Fmoc Strategies, Incorporation of Stable Pseudopeptide Bonds: Methylene Amino, Thioether, and Hydroxyethylene Derivatives, Synthesis of Conformationally Restricted Peptides, Purification of Synthetic Peptides by High Performance Liquid Chromatography, Molecular Weight Estimation for Neuropeptides Using Size-Exclusion High Performance Liquid Chromatography, Molecular Weight Determinations Using Polyacrylamide Gel Electrophoresis in Tris-Tricene Buffers, Determination of Neuropeptides by Capillary Electrophoresis, Characterization of Neuropeptide Processing by Fast Atom Bombardment Mass Spectrometry, Analysis of Neuropeptides by Size-Exclusion HPLC Linked to Electrospray Ionization Mass Spectrometry, Identification of Peptides by Matrix-Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF-MS) and Direct Analysis of the Laterobuccal Nerve from the Pond Snail Lymnaea stagnalis, Use of Circular Dichroism to Determine Secondary Structure of Neuropeptides, 1H-Nuclear Magnetic Resonance (NMR) in the Elucidation of Peptide Structure, The Study of Membrane- or Receptor-Bound Neuropeptides by NMR, Molecular Modeling of Neuropeptides, Tritium Labeling of Neuropeptides, The Use of IODO-GEN for Preparing 125I-Labeled Peptides and Their Purification by Reversed-Phase High Performance Liquid Chromatography, Production of Antisera Using Peptide Conjugates, Radioimmunoassay, Enzyme-Linked Immunosorbent Assay of Peptides, Sample Preparation for Peptide Immunocytochemistry, Immunocytochemical Methods for Regulatory Peptides, Ultrastructural Localization of Peptides Using Immunogold Labeling. Preparation of a Membrane Fraction for Receptor Studies and Solubilization of Receptor Proteins with Retention of Biological Activity, Radioligand Binding Using 125I-Labeled Peptides, Analysis of Data from "Cold Saturation" Radioligand Binding Experiments, Organ/Tissue Preparation for the Assessment of Agonist/ Antagonist Activity, Measurement of Efflux Rates from Brain to Blood, Assays of Neuropeptidases Using Fluorogenic Substrates, Characterization of Neuropeptides Using Inhibitors, Index.

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