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State University of New York at Stony Brook

6 ARTICLES PUBLISHED IN JoVE

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Biology

A Cell-to-cell Macromolecular Transport Assay in Planta Utilizing Biolistic Bombardment
Shoko Ueki 1, Benjamin L. Meyers 1, Farzana Yasmin 2, Vitaly Citovsky 2
1Department of Biochemistry and Cell Biology, State University of New York at Stony Brook, 2Bio-Medical Engineering Department, NED University of Engineering and Technology

Macromolecular trafficking between plant cells can be assessed by transiently expressing a fluorescently-tagged protein of interest and analyzing its intra- and intercellular distribution by confocal microscopy.

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Neuroscience

Studying the Integration of Adult-born Neurons
Yan Gu 1, Stephen Janoschka 1, Shaoyu Ge 1
1Department of Neurobiology & Behavior, State University of New York at Stony Brook

A way to study the integration of newborn dentate granule cells in adult animals is described. This technique uses an engineered retrovirus to label newborn neurons, followed by electrophysiological recordings to determine in vivo functional integration.

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Biology

Protein Membrane Overlay Assay: A Protocol to Test Interaction Between Soluble and Insoluble Proteins in vitro
Shoko Ueki 1, Benoît Lacroix 1, Vitaly Citovsky 1
1Department of Biochemistry and Cell Biology, State University of New York

Testing protein-protein interaction is indispensable for dissection of protein functionality. Here, we introduce an in vitro protein-protein binding assay to probe a membrane-immobilized protein with a soluble protein. This assay provides a reliable method to test interaction between an insoluble protein and a protein in solution.

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Immunology and Infection

Establishment of Epstein-Barr Virus Growth-transformed Lymphoblastoid Cell Lines
Joyce Hui-Yuen 1,2, Shane McAllister 1,2, Siva Koganti 2, Erik Hill 2, Sumita Bhaduri-McIntosh 1,2,3,4
1Stony Brook Children's Hospital, State University of New York at Stony Brook, 2Department of Pediatrics, State University of New York at Stony Brook, 3Department of Molecular Genetics, State University of New York at Stony Brook, 4Department of Microbiology, State University of New York at Stony Brook

We describe a method for generating transformed B cell lines using Epstein-Barr virus. We also illustrate a novel assay that can identify B cells destined to undergo transformation as early as three days after infection.

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JoVE Core

Protocol for the Synthesis of Ortho-trifluoromethoxylated Aniline Derivatives
Pengju Feng 1,2, Ming-Yu Ngai 1,2
1Department of Chemistry, State University of New York at Stony Brook, 2Institute of Chemical Biology and Drug Discovery, State University of New York at Stony Brook

An operationally simple procedure for the synthesis of ortho-trifluoromethoxylated aniline derivatives via a two-step sequence of O-trifluoromethylation of N-aryl-N-hydroxyacetamide followed by thermally induced intramolecular OCF3-migration is reported.

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Immunology and Infection

Optimization of a Quantitative Micro-neutralization Assay
Yipu Lin 1, Yan Gu 1, John W. McCauley 1
1Mill Hill Laboratory, The Francis Crick Institute

This study describes an imaging-based micro-neutralization assay to analyze the antigenic relationships between viruses. The protocol employs a flatbed scanner and has four steps, including titration, titration quantitation, neutralization, and neutralization quantitation. The assay works well with current circulating influenza A(H1N1)pdm09, A(H3N2), and B viruses.

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