Volume: 6 Issue: 5 —BIO-PROTOCOL

Cancer Biology

Mouse Mammary Intraductal (MIND) Method for Transplantation of Patient Derived Primary DCIS Cells and Cell Lines Authors: Frances Kittrell, Kelli Valdez, Hanan Elsarraj, Yan Hong, Daniel Medina and Fariba Behbod, date: 03/05/2016, view: 6711, Q&A: 0

The MIND method involves intraductal injection of patient derived ductal carcinoma in situ (DCIS) cells and DCIS cell lines (MCF10DCIS.COM and SUM225) inside the mouse mammary ducts [Video 1 and Figure 1 in Behbod et al. (2009)]. This method mimics the normal environment of DCIS and facilitates study of the natural progression of human ...

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Laser Microirradiation and Temporal Analysis of XRCC1 Recruitment to Single-strand DNA Breaks Authors: Anna Campalans and J. Pablo Radicella, date: 03/05/2016, view: 5961, Q&A: 0

The DNA molecule is exposed to a multitude of damaging agents that can compromise its integrity: single (SSB) and double strand breaks (DSB), intra- or inter-strand crosslinks, base loss or modification, etc. Many different DNA repair pathways coexist in the cell to ensure the stability of the DNA molecule. The nature of the DNA lesion ...

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Quantification of Rae-1 Staining Intensity in Glomeruli Authors: Roberto Spada and Domingo F. Barber, date: 03/05/2016, view: 4663, Q&A: 0

NKG2D is expressed on all NK cells and on subsets of NKT, CD8, CD4 and γδ T cells. NKG2D is activated by NKG2D ligands, a stress-induced family of MHC-I-like proteins. These ligands are upregulated on stressed/infected cells and are not widely expressed on healthy adult tissue. NKG2D ligands have been widely studied as potential targets for ...

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Immunology

Analysis of T Cell Proliferating and Polarizing Potential of Murine Dendritic Cells in Allogeneic-mixed Leukocyte Reaction Authors: Pawan Kumar and Sangeeta Bhaskar, date: 03/05/2016, view: 6956, Q&A: 0

Dendritic cells (DCs) play a critical role in mounting the T cell response against different infectious agents. Nature and intensity of the induced T cell responses are defined by activation status of DCs. It is generally accepted that IL-12, IL-4/IL-5 and IL-23 producing DCs induce T_H1, T_H2 and T_H17 type of immune ...

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Microbiology

Displacement-based ELISA: Quantifying Competition between Two Binding Partners for Interaction with a His-tagged Ligand Immobilized on a Ni²⁺-NTA Plate Authors: Allexa Dow and Sladjana Prisic, date: 03/05/2016, view: 6262, Q&A: 0

The displacement assay was designed to quantify the direct competition between two homologous ribosomal proteins from Mycobacterium tuberculosis, S18-1 and S18-2, for interaction with their cognate binding partner, ribosomal protein S6 (Prisic et al., 2015). The S18 proteins were dialyzed in two physiologically relevant ...

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Campylobacter jejuni γ-glutamyltranspeptidase Activity Assay Authors: Anne-Xander van der Stel and Marc M. S. M. Wösten, date: 03/05/2016, view: 5292, Q&A: 0

The enzyme γ-glutamyltranspeptidase (GGT, EC 2.3.2.2) is highly conserved among eukaryotic and prokaryotic organisms (Heisterkamp et al., 2008) and has a key function in glutathione metabolism. Although the enzyme is highly conserved and found throughout organisms ranging from bacteria to plants and animals several major difference ...

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Isolation of Genomic DNA from Mycobacterium Species Authors: Pawan Kumar, Soumitra Marathe and Sangeeta Bhaskar, date: 03/05/2016, view: 10978, Q&A: 1

Isolation of genomic DNA from Mycobacterium species has been a tedious procedure. This can primarily be attributed to thick and waxy cell wall of mycobacteria which hampers lysis of the bacterial cell. We have tested various approaches to isolate mycobacterial DNA and based on this, an optimized protocol is presented here. This protocol ...

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Testing the Effect of UV Radiation on the Survival of Burkholderia glumae Authors: Hari Sharan Karki and Jong Hyun Ham, date: 03/05/2016, view: 6925, Q&A: 0

Burkholderia glumae (B. glumae) is becoming a serious threat in the major rice producing areas of the world. It was reported that Burkholderia spp., including B. glumae, are adapted to a wide range of ecological niches. Different bacterial strains show different levels of UV tolerance which may be due to the ...

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Molecular Biology

Actin Retrograde Flow in Permeabilized Cells: Myosin-II Driven Centripetal Movement of Transverse Arcs Authors: Yee Han Tee and Alexander D. Bershadsky, date: 03/05/2016, view: 5808, Q&A: 0

Numerous biological functions such as cytokinesis, changes in cell shape and cell migration require actomyosin-driven cellular contractility. However, the detailed mechanism of how contractile forces drive cellular processes are difficult to decipher due to the complexity of the intracellular environment. In particular, the mesoscopic description ...

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Neuroscience

A Novel Task for Studying Memory of Occasional Events in Rats Authors: Marina Allerborn, Alexandra Gros, Belkacem Messaoudi, Damien Gervasoni, Samuel Garcia, Marc Thevenet, Serge Laroche, Alexandra Veyrac and Nadine Ravel, date: 03/05/2016, view: 5751, Q&A: 0

Episodic memory has been defined in humans as the conscious recollection of unique personal past experiences often occurring singly during daily life, including remembrance of what happened, where and when it happened (Tulving, 1972). Here, we propose and describe in details a novel protocol we recently used to test the ability of rats to form and ...

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Plant Science

Measurement of ³³P-PO₄ Absorption Capacity and Root-to-leaf Transfer in Arabidopsis Authors: Marie-Christine Thibaud and Elena Marin, date: 03/05/2016, view: 4764, Q&A: 0

This method allows quantification of phosphate absorption capacity by Arabidopsis roots using very simple equipment, and can be scaled up or down.

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Measurement of ³³P-PO₄ Absorption Kinetic Constants in Arabidopsis Authors: Elena Marin and Marie-Christine Thibaud, date: 03/05/2016, view: 4952, Q&A: 0

Based on the Michaelis-Menten kinetics model (Hofstee, 1952), this method allows calculation of the kinetic parameters (V_max, K_m) of phosphate uptake by Arabidopsis roots. This method is based on the quantification of phosphate uptake by Arabidopsis roots as described in Thibaud and Marin (2016), except that ...

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Immunogold Labeling Analysis of Cell Wall Polysaccharides with Special Reference to (1;3,1;4)-β-D-glucan in Rice Cell Walls Authors: Ryusuke Yokoyama, Natsumi Kido, Tsuyoshi Yamamoto, Jun Furukawa, Hiroaki Iwai, Shinobu Satoh and Kazuhiko Nishitani, date: 03/05/2016, view: 6301, Q&A: 0

Various types of cell wall compositions have evolved to fulfill a wide range of biological roles during the diversification of land plants. (1;3,1;4)-β-D-glucan (MLG) is a defining feature of the cell walls in the order Poales (Yokoyama and Nishitani, 2004), which has multiple functions associated with metabolic, growth, and defense systems. MLG ...

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Experimental Design to Determine Drought Stress Response and Early Leaf Senescence in Barley (Hordeum vulgare L.) Authors: Gwendolin Wehner, Christiane Balko and Frank Ordon, date: 03/05/2016, view: 11053, Q&A: 1

Premature leaf senescence induced by drought stress is a main factor for yield losses in barley. Research in drought stress tolerance has become more important as due to climate change the number of drought periods will increase and tolerance to drought stress has become a goal of high interest in barley breeding. However, reliable screening for ...

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Semi-thin Sectioning, Light and Fluorescence Microscopy of Floral Bud to Study Microspore Development in Arabidopsis Authors: Min-Jung Kim and Jungmook Kim, date: 03/05/2016, view: 6409, Q&A: 0

Pollen grains are male gametophytes produced within the pollen sacs of the anthers of the flower. Recent genetic studies have revealed several components involved in microspore development (Borg et al., 2009; Berger and Twell, 2011), and yet many components controlling microspore development remain to be identified. Semi-thin sectioning ...

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Shoot Regenerative Capacity Assays in Arabidopsis and Tobacco Authors: Tian-Qi Zhang and Jia-Wei Wang, date: 03/05/2016, view: 7321, Q&A: 0

Plant regeneration refers to a process through which an explant is differentiated to a whole plant under certain conditions. It has been shown that two plant hormones, auxin and cytokinin, play critical roles within this process (Duclercq et al., 2011). Cytokinin induces shoot regeneration, whereas auxin promotes root production. In ...

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Past Issues in 2016

Volume: 6 Issue: 5

Cancer Biology

Immunology

Microbiology

Molecular Biology

Neuroscience

Plant Science