Volume: 4 Issue: 4 —BIO-PROTOCOL

Biochemistry

Binding to Secreted Bone Matrix in vitro Authors: Aurélie Jeanne Tormo, Christian Beauséjour and Jean-François Gauchat, date: 02/20/2014, view: 6300, Q&A: 0

This method examines the bone matrix binding capacity of proteins. Using osteogenic differentiation medium, multipotent stromal cells (MSC) are induced to differentiate into osteocytes in vitro and to secrete bone matrix. The latter is confirmed using Alizarin red S staining, which detects the presence of calcific deposits ...

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A Surface Plasmon Resonance Method to Study HCV NS5B Inhibitors Authors: Melanie Wong and Giuseppe A. Papalia, date: 02/20/2014, view: 6162, Q&A: 0

Surface Plasmon Resonance (SPR) technology is a well-established platform used to evaluate the kinetic parameters of protein-small molecule interactions. Below, we describe the use of the ProteOn XPR36 biosensor from Bio-Rad (Hercules, CA) to evaluate the binding of small molecule inhibitors to recombinant NS5B protein. The high pI (> 9) of ...

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Cancer Biology

Isolation of Mammary Epithelial Cells and Fibroblasts From Mouse Tumor Author: Shiva Kazerounian, date: 02/20/2014, view: 14512, Q&A: 0

Developing cancer therapeutics requires the ability to investigate their effects using in vitro models of a specific type of tumor. This protocol provides a method for the isolation and adoption to growth in culture of cells from primary tumors. This is particularly valuable for studying mouse models where original tumor cells can be ...

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In vivo Extravasation Assay Authors: Elisa Penna and Daniela Taverna, date: 02/20/2014, view: 9382, Q&A: 1

Tumor metastases develop when disseminated intravascular cancer cells acquire the ability to arrest by adhering to the capillary walls of distant organs, actively extravasate into their parenchyma, proliferate and establish secondary colonies. The extravasation assay described here is an in vivo technique aimed to analyze the ability of ...

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Cell Biology

Lysosomal Amino Acid Efflux Assay Authors: Chunlei Cang and Dejian Ren, date: 02/20/2014, view: 6733, Q&A: 0

As the cellular “recycling” organelle, lysosomes break down proteins into amino acids, which are then transported into cytosol for reuse by various amino acid transporters. The transport rate of an amino acid is presumably regulated by cellular conditions such as organelle pH, membrane potential and metabolic states. Because of their ...

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Immunology

Isolation of Multipotent Stromal Cells from Mouse Bone Marrow Authors: Aurélie Jeanne Tormo, Moutih Rafei and Jean-François Gauchat, date: 02/20/2014, view: 6440, Q&A: 0

Generating mouse multipotent stromal cells (MSC) from bone-marrow cells is usefull for a wide range of applications. Effectively, these MSC can differentiate into adipocytes, osteocytes [See “Binding to Secreted Bone Matrix in vitro” (Tormo et al., 2014)] or ...

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DNA PCR Assays for Igh Rearrangement Authors: Yusuke Satoh, Takao Sudo and Takafumi Yokota, date: 02/20/2014, view: 6881, Q&A: 0

This protocol is used for the detection of immunoglobulin heavy (H) chain rearrangements. This PCR-based assay enables detection of D_H-J_H recombination in cultured hematopoietic cells (Schlissel et al., 1991; Satoh et al., 2013) [e.g. ES-derived cells (Satoh et al., 2013)].

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Chikungunya ELISA Protocol Authors: Caitlin Mattos Briggs, Amanda Piper and Raquel Hernandez, date: 02/20/2014, view: 6856, Q&A: 0

This protocol will result in the accurate qualitative measurement of anti-Chikungunya virus antibody (Ab) from infected mouse tissue or serum. This assay was developed by Dr. Caitlin Briggs, Arbovax, Inc. Chikungunya is a BL3 agent and should be handled in a biosafety level 3 laboratory under BL3 conditions. This protocol was used in the ...

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Isolation and Culture of Peritoneal Cell-derived Mast Cells Authors: Krisztina V. Vukman, Martin Metz, Marcus Maurer and Sandra M. O’Neill, date: 02/20/2014, view: 10406, Q&A: 2

The generation of mast cells for in vitro studies comes from a variety of sources including mast cell lines (MC/9) (McCurdy et al., 2001), bone marrow-derived mast cells (BMMCs) (Supajatura et al., 2001), skin-derived mast cells (FSMCs) (Matsushima et al., 2004), peritoneal-derived mast cells (PMCs) (Hochdorfer ...

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Isolation and Culture of Bone Marrow-derived Mast Cells Authors: Krisztina V. Vukman, Martin Metz, Marcus Maurer and Sandra M. O’Neill, date: 02/20/2014, view: 14302, Q&A: 5

The generation of mast cells for in vitro studies comes from a variety of sources including mast cell lines (MC/9) (McCurdy et al., 2001), bone marrow-derived mast cells (BMMCs) (Supajatura et al., 2001), skin-derived mast cells (FSMCs) (Matsushima et al., 2004), peritoneal-derived mast cells (PMCs) (Hochdorfer ...

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Microbiology

An ex vivo Model of HIV-1 Infection in Human Lymphoid Tissue and Cervico-vaginal Tissue Authors: Andrea Introini, Christophe Vanpouille, Jean Charles Grivel and Leonid Margolis, date: 02/20/2014, view: 10011, Q&A: 0

Human tissue explants are a valuable tool to study the interactions between host and infectious agents. They reliably mimic many important aspects of tissue cytoarchitecture and functions and allow us the investigation of the mechanisms of microbial pathogenesis under controlled laboratory conditions. One of the advantages of this system is that, ...

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Stem Cell

Limiting Dilution Assays to Determine Frequencies of Lymphohematopoietic Progenitors Authors: Takafumi Yokota and Yusuke Satoh, date: 02/20/2014, view: 7191, Q&A: 0

This protocol is useful to determine the frequencies of lymphohematopoietic progenitors in tested samples. To effectively support the growth and differentiation of primitive lymphohematopoietic progenitors, complex signals from stromal cells are important. Several stromal cell lines are known to support both lymphoid and myeloid cells ...

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Past Issues in 2014

Volume: 4 Issue: 4

Biochemistry

Cancer Biology

Cell Biology

Immunology

Microbiology

Stem Cell