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Latest Articles

Basic Techniques in Mammalian Cell Tissue Culture
Katy Phelan, Kristin M. May

Cultured mammalian cells are used extensively in cell biology studies. It requires a number of special skills in order to be able to preserve the structure, function, behavior, and biology of the cells in culture.

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Isolation and Culture of Spinal Cord Motor Neurons
Alice Klausmeyer, D. Stern, S. Wiese

Isolated spinal motoneurons are a powerful tool for studying basic mechanisms of neurite growth and survival. Since motoneurons are a minor population of developing spinal cord cells, they need to be purified and enriched to separate them from non‐neuronal cells.

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Isolation of Integrin‐Based Adhesion Complexes
Matthew C. Jones, Jonathan D. Humphries, Adam Byron, Angélique Millon‐Frémillon, Joseph Robertson, Nikki R. Paul, Daniel H. J. Ng, Janet A. Askari, Martin J. Humphries

The integration of cells with their extracellular environment is facilitated by cell surface adhesion receptors, such as integrins, which play important roles in both normal development and the onset of pathologies.

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Adhesive Micropatterns to Study Intermediate Filament Function in Nuclear Positioning
Isabelle Dupin, Julien Elric, Sandrine Etienne‐Manneville

The nucleus is generally found near the cell center; however its position can vary in response to extracellular or intracellular signals, leading to a polarized intracellular organization. Nuclear movement is mediated by the cytoskeleton and its associated motors.

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Reconstitution of a Bioengineered Salivary Gland Using a Three‐Dimensional Cell Manipulation Method
Miho Ogawa, Takashi Tsuji

One concept in regenerative therapy is the replacement of a lost or damaged organ with a regenerated, fully functional organ.

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Diffraction‐Unlimited Fluorescence Microscopy of Living Biological Samples Using pcSOFI
Sam Duwé, Benjamien Moeyaert, Peter Dedecker

The complex microscopic nature of many live biological processes is often obscured by the diffraction limit of light, requiring diffraction‐unlimited fluorescence microscopy to resolve them.

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Preparation and Separation of DNA‐Wrapped Carbon Nanotubes
Geyou Ao, Ming Zheng

Purification of single‐chirality single‐wall carbon nanotubes (SWCNTs) from their synthetic mixture is a prerequisite for many applications.

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SNIPER Peptide‐Mediated Degradation of Endogenous Proteins
Xuelai Fan, Yu Tian Wang

Rapid and reversible methods for altering the function of endogenous proteins are not only indispensable tools for probing complex biological systems, but may potentially drive the development of new therapeutics for the treatment of human diseases.

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Universal Non‐Antibody Detection of Protein Phosphorylation Using pIMAGO
Anton B. Iliuk, W. Andy Tao

This article describes methods for a new, non‐antibody phosphorylation detection reagent, termed pIMAGO (phospho‐imaging).

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Glucose Tolerance Tests for Systematic Screening of Glucose Homeostasis in Mice
Jan Rozman, Birgit Rathkolb, Susanne Neschen, Helmut Fuchs, Valérie Gailus‐Durner, Martin Klingenspor, Eckhard Wolf, Martin Hrabě de Angelis

This article presents a detailed description of intraperitoneal and oral glucose tolerance tests in mice. The former is widely used in initial high‐throughput phenotyping of mutant mice to assess a diabetic phenotype and alterations in glucose homeostasis.

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Quantitative Proteomic Approaches in Mouse: Stable Isotope Incorporation by Metabolic (SILAC) or Chemical Labeling (Reductive Dimethylation) Combined with High‐Resolution Mass Spectrometry
Anja M. Billing, Hisham Ben Hamidane, Johannes Graumann

Mass spectrometry–based quantitative proteomics is a powerful method for in‐depth exploration of protein expression, allowing researchers to probe its regulation and study signal‐transduction networks, protein turnover, secretion, and spatial distribution, as well as post‐translational modi.

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Mouse Models of Bone Healing: Fracture, Marrow Ablation, and Distraction Osteogenesis
Kyle Lybrand, Beth Bragdon, Louis Gerstenfeld

Three commonly used murine surgical models of bone healing [closed fracture with intramedullary fixation, distraction osteogenesis (DO), and marrow ablation by reaming] are presented. Detailed surgical protocols for each model are outlined.

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Overview of Biological Mechanisms and Applications of Three Murine Models of Bone Repair: Closed Fracture with Intramedullary Fixation, Distraction Osteogenesis, and Marrow Ablation by Reaming
Beth Bragdon, Kyle Lybrand, Louis Gerstenfeld

Fractures are one of the most common large‐organ, traumatic injuries in humans, and osteoporosis‐related fractures are the fastest growing health care problem of aging.

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Mouse Anesthesia and Analgesia
Sean Adams, Cholawat Pacharinsak

Providing anesthesia and analgesia for mouse subjects is a common and critical practice in the laboratory setting.

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Arrestin‐3‐Dependent Activation of c‐Jun N‐Terminal Kinases (JNKs)
Xuanzhi Zhan, Seunghyi Kook, Tamer S. Kaoud, Kevin N. Dalby, Eugenia V. Gurevich, Vsevolod V. Gurevich

Only one out of four mammalian arrestin subtypes, arrestin‐3, facilitates the activation of JNK family kinases. Here we describe two different protocols used for elucidating the mechanisms involved.

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Single‐Channel Recording of Glutamate Receptors
Chris Shelley

Highlighted in this unit are issues that should be considered when recording glutamate receptors at the single‐channel level, including some commonly encountered problems and their remedies. “UNIT 11.

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Single‐Channel Analysis of Glutamate Receptors
Chris Shelley

This is a companion to UNIT 11.16: Single‐Channel Recording of Glutamate Receptors. Described here are techniques for analyzing single‐channel currents recorded from glutamate receptors to characterize their properties.

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Generation of Human Acute Lymphoblastic Leukemia Xenografts for Use in Oncology Drug Discovery
Linda Holmfeldt, Charles G. Mullighan

The establishment of reproducible mouse models of acute lymphoblastic leukemia (ALL) is necessary to provide in vivo therapeutic test systems that recapitulate human ALL, and for amplification of limited amounts of primary tumor material.

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