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Determination of Molecular Size by Size‐Exclusion Chromatography (Gel Filtration)

Abstract Table of Contents Materials Figures Literature Cited Abstract Size?exclusion or gel filtration chromatography is one of the most popular methods for determining the sizes of proteins. Proteins in

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Determination of Molecular Size by Zonal Sedimentation Analysis on Sucrose Density Gradients

Abstract Table of Contents Materials Figures Literature Cited Abstract The molecular weight of a protein is a basic characteristic that can only be approximated by techniques such as gel filtration and el

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High‐Throughput Screening for Protein‐Protein Interactions Using Yeast Two‐Hybrid Arrays

Abstract Table of Contents Materials Figures Literature Cited Abstract Arrays are used for parallel assays of large numbers of proteins in a single experiment. They provide an alternative to libraries for

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Modification of the 5′ Terminus of Oligodeoxyribonucleotides for Conjugation with Ligands

Abstract Table of Contents Materials Figures Literature Cited Abstract Ligands can be introduced at the 5? terminus of an oligonucleotide by adding a linker to the ligand and modifying the 5? terminus of

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Incorporation of Halogenoalkyl, 2‐Pyridyldithioalkyl, or Isothiocyanate Linkers into Ligands

Abstract Table of Contents Materials Figures Literature Cited Abstract Ligands can be introduced at the 5' terminus of an oligonucleotide by adding a linker to the ligand and modifying the 5' terminus of

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Size‐Exclusion Chromatography with On‐Line Light Scattering

Abstract Table of Contents Materials Figures Literature Cited Abstract This unit describes the use of size?exclusion chromatography with on?line light scattering, UV absorbance, and refractive index detec

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Identification of Protein Interactions by Far Western Analysis

Abstract Table of Contents Materials Figures Literature Cited Abstract This unit describes far western blotting, a method of identifying protein?protein interactions. In a far western blot, one protein of

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Characterization of Calcium Channel Binding

Abstract Table of Contents Materials Figures Literature Cited Abstract Voltage?dependent calcium channels are expressed in a variety of tissues including heart, muscles and brain. Saturation binding of a

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Characterization of Chemokine Receptors

Abstract Table of Contents Materials Figures Literature Cited Abstract This unit describes the procedures for measuring binding of a radiolabeled chemokine to chemokine receptors in cells or cell membrane

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Metabolic Labeling and Immunoprecipitation of Drosophila Proteins

Abstract Table of Contents Materials Literature Cited Abstract The genetics of Drosophila is a powerful tool in the analysis of mutants and mutant proteins. Cultures of cells derived from wild?type or mutant flies can be pulse labeled to b

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Conjugation of 5′‐Functionalized Oligodeoxyribonucleotides with Properly Functionalized Ligands

Abstract Table of Contents Materials Figures Literature Cited Abstract This unit reports the synthesis of oligodeoxyribonucleotides covalently linked via their 5? termini to various ligands such as interc

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Metabolic Labeling of Glycoproteins with Radioactive Sugars

Abstract Table of Contents Materials Figures Literature Cited Abstract This unit describes methods for preparation of glycoproteins metabolically labeled with radioactive sugars, sulfate, and phosphate. M

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Scintillation Proximity Assay (SPA) Technology to Study Biomolecular Interactions

Abstract Table of Contents Materials Figures Literature Cited Abstract Scintillation proximity assay (SPA) is a versatile homogeneous technique for radioactive assays which eliminates the need for separat

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Chromatin Assembly Using Drosophila Systems

Abstract Table of Contents Materials Figures Literature Cited Abstract To successfully study chromatin structure and activity in vitro, it is essential to have a chromatin assembly system that will prepar

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Analysis of Oxidative Modification of Proteins

Abstract Table of Contents Materials Figures Literature Cited Abstract Reactions between protein molecules and reactive oxygen species (ROS) often lead to the modification of certain amino acid residues s

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Radioligand Binding Assays for the Glycine Site on N‐Methyl‐D‐Aspartate Receptors

Abstract Table of Contents Materials Figures Literature Cited Abstract This unit describes a competitive binding assay for the glycine binding site on the NMDA subtype of glutamate receptors in rat brain

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Preparation of Gold Nanoparticle–DNA Conjugates

Abstract Table of Contents Materials Figures Literature Cited Abstract This unit describes the preparation of conjugates between nanometer?scale gold particles and synthetic oligonucleotides. Oligonucleotid

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Radioiodination of Cellular Proteins

Abstract Table of Contents Materials Figures Literature Cited Abstract Despite the advances of non?radioactive methods of protein labeling, radioiodination remains an important tool for studying proteins.

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Techniques for Measuring the Activity of Carboxylic Acid:CoA Ligase and Acyl‐CoA:Amino Acid N‐Acyltransferase: The Amino Acid Conjugation Pathway

Abstract Table of Contents Materials Figures Literature Cited Abstract A wide variety of xenobiotic carboxylic acids are metabolized to their amino acid conjugates via a pathway that exists primarily in l

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Synthesis, Characterization, and Application of Substituted Pyrazolopyrimidine Nucleosides

Abstract Table of Contents Materials Figures Literature Cited Abstract This unit describes, in detail, the preparation of 3?aminopropyl?substituted pyrazolopyrimidine analogs of the purines deoxyadenosine

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