• Volume 12, 2022
  • Volume 11, 2021
  • Volume 10, 2020
  • Volume 9, 2019
  • Volume 8, 2018
  • Volume 7, 2017
  • Volume 6, 2016
  • Volume 5, 2015
  • Volume 4, 2014
  • Volume 3, 2013
  • Volume 2, 2012
  • Volume 1, 2011


Extraction and Profiling of Plant Polar Glycerol Lipids Authors:  Yu Liu and Xuemin Wang, date: 06/20/2016, view: 10571, Q&A: 0
This protocol describes a method to extract total polar glycerol lipids from plant materials, followed by mass spectrometry profiling. Different glycerol lipid classes can be distinguished by their head-groups, which can be profiled automatically and quantitatively by a triple quadrupole mass spectrometry in multiple reaction monitoring (MRM) mode ...
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Positional Analysis of Fatty Acids in Phospholipids by PLA2 Treatment Authors:  Yu Liu and Xuemin Wang, date: 06/20/2016, view: 6598, Q&A: 0
Plant phospholipids can be produced in the endoplasmic reticulum or plastids. Lipids from different sources can be distinguished by the fatty acid profile, in terms of the preferred fatty acid species esterified to the sn-1 or sn-2 position of the glycerol backbone (Ohlrogge and Browse, 1995). This protocol is used to determine ...
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Cell Biology

Protocol for Microfluidic System to Automate the Preparation and Fractionation of the Nucleic Acids in the Cytoplasm Versus Nuclei of Single Cells Authors:  Kentaro Kuriyama, Hirofumi Shintaku and Juan G. Santiago, date: 06/20/2016, view: 7719, Q&A: 0
This protocol describes the extraction, fractionation, and recovery of cytoplasmic nucleic acids (e.g., cytoplasmic RNA) versus nucleic acids in the cell nucleus (including genomic DNA, gDNA) from single cells with a microfluidic system. The method enables independent, sequence-specific analyses of these critical markers (Kuriyama et al. ...
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Whole-mount Enteroid Proliferation Staining Authors:  Caitlyn W. Barrett, Sarah P. Short, Yash A. Choksi and Christopher Shawn Williams, date: 06/20/2016, view: 10546, Q&A: 0
Small intestinal organoids, otherwise known as enteroids, have become an increasingly utilized model for intestinal biology in vitro as they recapitulate the various epithelial cells within the intestinal crypt (Mahe et al., 2013; Sato et al., 2009). Assessment of growth dynamics within these cultures is an important ...
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In vivo OVA-specific Cytotoxic CD8+ T Cell Killing Assay Authors:  Nada Chaoul, Catherine Fayolle and Claude Leclerc, date: 06/20/2016, view: 18254, Q&A: 0
Cytotoxic CD8+ T cells are responsible for the lysis of cells expressing peptides associated with MHC class I molecules and derived from infection with a pathogen or from mutated antigens. In order to quantify in vivo this antigen-specific CD8+ T cell killing activity, we use the in vivo killing assay (IVK). ...
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Measurement of Mitochondrial DNA Release in Response to ER Stress Authors:  Denise N. Bronner and Mary X. O’Riordan, date: 06/20/2016, view: 15181, Q&A: 0
Mitochondria house the metabolic machinery for cellular ATP production. The mitochondrial network is sensitive to perturbations (e.g., oxidative stress and pathogen invasion) that can alter membrane potential, thereby compromising function. Healthy mitochondria maintain high membrane potential due to oxidative phosphorylation (Ly ...
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Isolation and Culture of the Islets of Langerhans from Mouse Pancreas Authors:  Kate L Graham, Stacey Fynch, Evan G Pappas, Christina Tan, Thomas WH Kay and Helen E Thomas, date: 06/20/2016, view: 22716, Q&A: 1
The islets of Langerhans are clusters of endocrine cells located within the pancreas. Insulin-producing beta cells are the major cell type within islets, with glucagon-producing alpha cells and somatostatin-producing delta cells the other major cell types. The beta cells are the target of immune-mediated destruction in type 1 diabetes (Graham et ...
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Efficient Isolation of Influenza Specific CTLs Authors:  Prerak Trivedi and Stuart I. Mannering, date: 06/20/2016, view: 7895, Q&A: 0
Human antigen-specific CD8+ T-cell clones are valuable tools for dissecting CD8+ T-cell responses against antigens derived from infectious agents, cancer and self antigens. Here we describe a protocol for isolating human antigen-specific CD8+ T cells. This protocol uses surface capture of IFNγ to identify antigen ...
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Aspergillus terreus Infection of Fruits and Terrein Quantification by HPLC Analysis Authors:  Markus Gressler and Matthias Brock, date: 06/20/2016, view: 7631, Q&A: 0
The opportunistic fungal human and plant pathogen Aspergillus terreus (A. terreus) can be isolated from sea water, soil or decaying organic matter such as rotting leaves and fruits. While growing on fruits A. terreus produces secondary metabolites such as terrein, which may ease its penetration into plant tissues. In ...
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Molecular Biology

Micro-chromatin Immunoprecipitation (μChIP) Protocol for Real-time PCR Analysis of a Limited Amount of Cells Authors:  Sébastien Gillotin and François Guillemot, date: 06/20/2016, view: 8314, Q&A: 0
Chromatin immunoprecipitation followed by deep sequencing (ChIP-Seq) is an important strategy to study gene regulation. When availability of cells is limited, however, it can be useful to focus on specific genes to investigate in depth the role of transcription factors or histone marks. Unfortunately, performing ChIP experiments to study ...
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Plant Science

Cytology and Microscopy: Immunolocalization of Covalently Modified Histone Marks on Barley Mitotic Chromosomes Authors:  Isabelle Colas, Katie Baker and Andrew J. Flavell, date: 06/20/2016, view: 8319, Q&A: 0
Barley is a diploid inbreeding crop with a genome of 5 GB organized into seven chromosomes. The relatively low chromosome number and their large size make barley an excellent model for chromosome cytogenetic studies of large genome cereal crops. Chromatin can be defined as euchromatin or heterochromatin. Euchromatin is gene-rich, less condensed, ...
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Measuring Auxin Transport Capacity in Seedling Roots of Medicago truncatula Authors:  Jason Liang Pin Ng and Ulrike Mathesius, date: 06/20/2016, view: 8688, Q&A: 0
Measurement of auxin transport capacity provides quantitative data on the physiological machinery involved in auxin transport within plants. This technique is easy to perform and gives quick results. Radiolabelled auxin (indole-3-acetic-acid) is fed into the roots of Medicago truncatula via an agar block. The resulting radioactivity from ...
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Quantifying Auxin Metabolites in Young Root Tissue of Medicago truncatula by Liquid Chromatography Electrospray-ionisation Quadrupole Time-of-flight (LC-ESI-QTOF) Tandem Mass Spectrometry Authors:  Jason Liang Pin Ng, Thy T. Truong, Charles H. Hocart and Ulrike Mathesius, date: 06/20/2016, view: 9015, Q&A: 0
Auxins represent a major group of phytohormones controlling plant development. The spatio-temporal regulation of auxin gradients is essential for the initiation, growth and correct development of plant organs. Because auxins and their metabolites occur at trace levels in plant tissue, experiments requiring identification plus their selective and ...
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Root-knot Nematode Penetration and Sclareol Nematicidal Activity Assays Authors:  Taketo Fujimoto, Takayuki Mizukubo, Hiroshi Abe and Shigemi Seo, date: 06/20/2016, view: 9460, Q&A: 0
Plant parasitic nematodes parasitize roots and/or stems of various plants thereby inhibiting absorption of nutrients and moisture. In particular, root-knot nematodes (RKN) are a group of the most devastating pests. Various techniques, such as soil sterilization, cultivation of resistant crops, and chemical application, have been developed to ...
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Stem Cell

Pit Assay to Measure the Bone Resorptive Activity of Bone Marrow-derived Osteoclasts Authors:  Alexander Vesprey and Wentian Yang, date: 06/20/2016, view: 12654, Q&A: 0
Although it is possible to use a tartrate-resistant acid phosphatase (TRAP) stain to assist in identifying osteoclasts, a separate method is needed to determine the bone resorption activity of osteoclasts. Since osteoclasts leave “pits” after bone matrix resorption (Charles et al., 2014), it is possible to stain pits as a method of ...
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