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Purification of Protein-complexes from the Cyanobacterium Synechocystis sp. PCC 6803 Using FLAG-affinity Chromatography

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Spectrophotometric Assessment of Heme Oxygenase-1 Activity in Leishmania-infected Macrophages

Authors: Moumita Basu, Shriya Saha and Anindita Ukil, date: 04/05/2020, view: 368, Q&A: 0

[Abstract] Heme oxygenase-1 (HO-1) is a stress responsive enzyme that metabolizes heme and releases free iron, carbon monoxide (CO), and biliverdin (BV), which rapidly undergoes conversion to bilirubin (BL). Estimation of bilirubin is the basis of HO-1 assay. HO-1 activity is widely employed to determine antioxidant response of cells under different ...

Cell Microencapsulation and Cryopreservation with Low Molecular Weight Hyaluronan and Dimethyl Sulfoxide

Authors: H. Gurruchaga, L. Saenz del Burgo, G. Orive, R. M. Hernandez, J. Ciriza and J. L. Pedraz, date: 02/20/2019, view: 1945, Q&A: 0

[Abstract] Cryopreservation is commonly used for the storage of cells, tissues, organs or 3D cell-based products using ultra-low temperatures, which involves the immersion in liquid nitrogen for their long-term preservation. The cryopreservation of several microencapsulated cells is usually performed by the slow freezing with the dimethyl sulfoxide (DMSO) as ...

Murine Pharmacokinetic Studies

Authors: Alix F. Leblanc, Kevin M. Huang, Muhammad Erfan Uddin, Jason T. Anderson, Mingqing Chen and Shuiying Hu, date: 10/20/2018, view: 5497, Q&A: 0

[Abstract] Murine pharmacokinetics (PK) represents the absorption, distribution, metabolism, and elimination of drugs from the body, which helps to guide clinical studies, ultimately resulting in more effective drug treatment. The purpose of this protocol is to describe a serial bleeding protocol, obtaining blood samples at six time points from single mouse ...

Using Stable Isotopes in Bone Marrow Derived Macrophage to Analyze Metabolism

Authors: Chih-Wei Ko, Daniel Counihan, David DeSantis, Zach Sedor-Schiffhauer, Michelle Puchowicz and Colleen M Croniger, date: 09/05/2018, view: 2684, Q&A: 0

[Abstract] Using gas chromatography mass spectrometry (GC-MS) to analyze the citric acid cycle (CAC) and related intermediates (such as glutamate, glutamine, GABA, and aspartate) is an analytical approach to identify unexpected correlations between apparently related and unrelated pathways of energy metabolism. Intermediates can be as expressed as their ...

In vivo and in vitro ³¹P-NMR Study of the Phosphate Transport and Polyphosphate Metabolism in Hebeloma cylindrosporum in Response to Plant Roots Signals

Authors: Christine Le Guernevé, Adeline Becquer, Margarita Torres-Aquino, Laurie K Amenc, Carlos Trives-Segura, Siobhan Staunton, Claude Plassard and Hervé Quiquampoix, date: 08/20/2018, view: 2659, Q&A: 0

[Abstract] We used in vivo and in vitro phosphorus-31 nuclear magnetic resonance (³¹P-NMR) spectroscopy to follow the change in transport, compartmentation and metabolism of phosphate in the ectomycorrhizal fungus Hebeloma cylindrosporum in response to root signals originating from host (Pinus pinaster) or ...

Measurement of Oxygen Consumption Rate (OCR) and Extracellular Acidification Rate (ECAR) in Culture Cells for Assessment of the Energy Metabolism

Authors: Birte Plitzko and Sandra Loesgen, date: 05/20/2018, view: 37027, Q&A: 0

[Abstract] Mammalian cells generate ATP by mitochondrial (oxidative phosphorylation) and non-mitochondrial (glycolysis) metabolism. Cancer cells are known to reprogram their metabolism using different strategies to meet energetic and anabolic needs (Koppenol et al., 2011; Zheng, 2012). Additionally, each cancer tissue has its own individual ...

Measurement of ROS in Caenorhabditis elegans Using a Reduced Form of Fluorescein

Authors: Shaarika Sarasija and Kenneth R. Norman, date: 04/05/2018, view: 3185, Q&A: 0

[Abstract] Oxidative stress is implicated in the pathogenesis of various neurodegenerative diseases, including Alzheimer’s disease. Oxidative stress is a result of a disruption of the equilibrium between antioxidants and oxidants, in favor of oxidants. Since mitochondria are major sites of production and reduction of reactive oxygen species (ROS), ...

GC/MS-based Analysis of Volatile Metabolic Profile Along in vitro Differentiation of Human Induced Pluripotent Stem Cells

Authors: Rosamaria Capuano, Rosa Valentina Talarico, Paola Spitalieri, Roberto Paolesse, Giuseppe Novelli, Federica Sangiuolo and Corrado Di Natale, date: 12/05/2017, view: 5779, Q&A: 0

[Abstract] Human induced pluripotent stem cells (hiPSCs) are a promising tool in cell-based therapies for degenerative diseases. A safe application of hiPSCs in vivo, requires the detection of the presence of residual undifferentiated pluripotent cells that can potentially cause the insurgence of teratomas. Several studies point out that metabolic ...

A Simple and Rapid Assay for Measuring Phytoalexin Pisatin, an Indicator of Plant Defense Response in Pea (Pisum sativum L.)

Authors: Lee A. Hadwiger and Kiwamu Tanaka, date: 07/05/2017, view: 4539, Q&A: 0

[Abstract] Phytoalexins are antimicrobial substance synthesized in plants upon pathogen infection. Pisatin (Pisum sativum phytoalexin) is the major phytoalexin in pea, while it is also a valuable indicator of plant defense response. Pisatin can be quantitated in various methods from classical organic chemistry to ...

Simple Spectroscopic Determination of Nitrate, Nitrite, and Ammonium in Arabidopsis thaliana

Authors: Takushi Hachiya and Yuki Okamoto, date: 05/20/2017, view: 11189, Q&A: 1

[Abstract] Plants use nitrate, nitrite, and ammonium as inorganic nitrogen (N) sources. These N compounds are included in plant tissues at various concentrations depending on the balance between their uptake and assimilation. Thus, the contents of nitrate, nitrite, and ammonium are physiological indicators of plant N economy. Here, we describe a protocol for ...

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