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Proteolytic Fingerprinting of Complex Biological Samples Using Combinatorial Libraries of Fluorogenic Probes

Abstract Table of Contents Materials Figures Literature Cited Abstract Proteases have garnered interest as candidate biomarkers and therapeutic targets for many human diseases. A key challenge is the iden

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Fluorescent Labeling of Specific Cysteine Residues Using CyMPL

Abstract Table of Contents Materials Figures Literature Cited Abstract The unique reactivity and relative rarity of cysteine among amino acids makes it a convenient target for the site?specific chemical m

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Synthesis of Oligodeoxynucleotides with 5′‐Caps Binding RNA Targets

Abstract Table of Contents Materials Figures Literature Cited Abstract Protocols for the synthesis of oligodeoxynucleotides with a short peptidyl substituent linked to the 5??O?terminus through a phosphodie

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Specificity Profiling of Protein‐Binding Domains Using One‐Bead‐One‐Compound Peptide Libraries

Abstract Table of Contents Materials Figures Literature Cited Abstract One?bead?one?compound (OBOC) libraries consist of structurally related compounds (e.g., peptides) covalently attached to a solid supp

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A High‐Throughput Screening Method for Identification of Inhibitors of the Deubiquitinating Enzyme USP14

Abstract Table of Contents Materials Figures Literature Cited Abstract Deubiquitinating enzymes (DUBs) reverse the process of ubiquitination, and number nearly 100 in humans. In principle, DUBs represent

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Synchronizing Protein Transport in the Secretory Pathway

Abstract Table of Contents Materials Figures Literature Cited Abstract To be secreted or transported to their target compartments, newly synthesized proteins leave the endoplasmic reticulum to reach the G

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Visualization of Live Primary Cilia Dynamics Using Fluorescence Microscopy

Abstract Table of Contents Materials Figures Videos Literature Cited Abstract Methods useful for exploring the formation and functions of primary cilia in living cells are

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Using CellX to Quantify Intracellular Events

Abstract Table of Contents Figures Literature Cited Abstract Methods to quantify features of individual cells using light microscopy have become widely used in biology. A multitude of computational tools has been developed for image analys

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Overview of Current Methods in Sedimentation Velocity and Sedimentation Equilibrium Analytical Ultracentrifugation

Abstract Table of Contents Figures Literature Cited Abstract Modern computational strategies have allowed for the direct modeling of the sedimentation process of heterogeneous mixtures, resulting in sedimentation velocity (SV) size?distrib

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Overview of Determination of Biopharmaceutical Properties for Development Candidate Selection

Abstract Table of Contents Literature Cited Abstract The physicochemical and biopharmaceutical properties of putative drug molecules impact their performance in both in vitro and in vivo studies. The design and selection of molecules with drug?like properties assists in the

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Biotinylation of a Propargylated Cyclic (3′‐5′) Diguanylic Acid and of Its Mono‐6‐Thioated Analog Under “Click” Conditions

Abstract Table of Contents Materials Figures Literature Cited Abstract Commercial N2?isobutyryl?5??O?(4,4??dimethoxytrityl)?2??O?(propargyl)guanosine is converted to its 3??O?levulinyl ester in a yield of 9

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Use of a Novel 5′‐Regioselective Phosphitylating Reagent for One‐Pot Synthesis of Nucleoside 5′‐Triphosphates from Unprotected Nucleosides

Abstract Table of Contents Materials Figures Literature Cited Abstract 5??Triphosphates are building blocks for enzymatic synthesis of DNA and RNA. This unit presents a protocol for convenient synthesis o

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Detecting Low‐Affinity Extracellular Protein Interactions Using Protein Microarrays

Abstract Table of Contents Materials Figures Literature Cited Abstract Low?affinity extracellular protein interactions are critical for cellular recognition processes, but are not generally detected by me

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Preparation of Recombinant Protein Spotted Arrays for Proteome‐Wide Identification of Kinase Targets

Abstract Table of Contents Materials Figures Literature Cited Abstract Protein microarrays allow unique approaches for interrogating global protein interaction networks. Protein arrays can be divided into

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Protein Microarrays for Identification of Novel Extracellular Protein‐Protein Interactions

Abstract Table of Contents Materials Figures Literature Cited Abstract Functional protein microarrays offer the capability for high?throughput protein interaction analysis and have long promised to be a p

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Overview of Protein Microarrays

Abstract Table of Contents Figures Literature Cited Abstract Protein microarray technology is an emerging field that provides a versatile platform for the characterization of hundreds of thousands of proteins in a highly parallel and high?

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A Detailed Protocol for Formaldehyde‐Assisted Isolation of Regulatory Elements (FAIRE)

Abstract Table of Contents Materials Figures Literature Cited Abstract Nucleosome displacement is a key event in the regulation of gene expression in the eukaryotic genome. This unit details an approach c

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Preparation of Oligodeoxyribonucleotides Containing the Pyrimidine(6–4)pyrimidone Photoproduct by Using a Dinucleotide Building Block

Abstract Table of Contents Materials Figures Literature Cited Abstract This unit describes procedures for the synthesis of a dinucleotide?type building block of the pyrimidine(6?4)pyrimidone photoproduct

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Site‐Specific Protein Labeling with SNAP‐Tags

Abstract Table of Contents Materials Figures Literature Cited Abstract Site?specific labeling of cellular proteins with chemical probes is a powerful tool for studying protein function in living cells. A

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Characterization of P1 (Adenosine) Purinoceptors

Abstract Table of Contents Materials Figures Literature Cited Abstract The purine nucleoside adenosine (ADO) is an important modulator of cellular function in mammalian tissues, modulating cellular functi

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