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Hypochlorous Acid Staining with R19-S in the Drosophila Intestine upon Ingestion of Opportunistic Bacteria

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Quantification of the Humidity Effect on HR by Ion Leakage Assay

Authors: Musoki Mwimba and Xinnian Dong, date: 04/05/2019, view: 407, Q&A: 0

[Abstract] We describe a protocol to measure the contribution of humidity on cell death during the effector-triggered immunity (ETI), the plant immune response triggered by the recognition of pathogen effectors by plant resistance genes. This protocol quantifies tissue cell death by measuring ion leakage due to loss of membrane integrity during the ...

Soluble and Solid Iron Reduction Assays with Desulfitobacterium hafniense

Authors: Lucrezia Comensoli, Julien Maillard, Wafa M. Kooli, Pilar Junier and Edith Joseph, date: 09/05/2018, view: 1033, Q&A: 0

[Abstract] There is a pressing need to develop sustainable and efficient methods to protect and stabilize iron objects. To develop a conservation-restoration method for corroded iron objects, this bio-protocol presents the steps to investigate reductive dissolution of ferric iron and biogenic production of stabilizing ferrous iron minerals in the strict ...

Evaluation of Root pH Change Through Gel Containing pH-sensitive Indicator Bromocresol Purple

Authors: Aparecida L. Silva, Keini Dressano, Paulo H. O. Ceciliato, Juan Carlos Guerrero-Abad and Daniel S. Moura, date: 04/05/2018, view: 2350, Q&A: 0

[Abstract] The Rapid Alkalinization Factor (RALF) is a plant hormone peptide that inhibits proton transport causing alkalinization of the extracellular media. To detect the alkalinization response elicited by RALF peptides in root cells, Arabidopsis seedlings are carefully transferred to a gel containing the pH-sensitive indicator bromocresol ...

Quantification of Plant Cell Death by Electrolyte Leakage Assay

Authors: Noriyuki Hatsugai and Fumiaki Katagiri, date: 03/05/2018, view: 5566, Q&A: 1

[Abstract] We describe a protocol to measure the electrolyte leakage from plant tissues, resulting from loss of cell membrane integrity, which is a common definition of cell death. This simple protocol is designed to measure the electrolyte leakage from a tissue sample over a time course, so that the extent of cell death in the tissue can be monitored ...

Extraction and Molybdenum Blue-based Quantification of Total Phosphate and Polyphosphate in Parachlorella

Authors: Shuhei Ota and Shigeyuki Kawano, date: 09/05/2017, view: 4066, Q&A: 0

[Abstract] Inorganic phosphorus is a non-renewable resource and an essential element for life on Earth. Organisms such as algae, protists, and animals can store phosphate (Pi) through uptake of Pi as polyphosphate (poly-P), which is a linear polymer of orthophosphate residues linked by high-energy phosphoanhydride bonds. Here, we describe procedures for ...

Bacterial Intracellular Sodium Ion Measurement using CoroNa Green

Authors: Yusuke V. Morimoto, Keiichi Namba and Tohru Minamino, date: 01/05/2017, view: 4962, Q&A: 0

[Abstract] The bacterial flagellar type III export apparatus consists of a cytoplasmic ATPase complex and a transmembrane export gate complex, which are powered by ATP and proton motive force (PMF) across the cytoplasmic membrane, respectively, and transports flagellar component proteins from the cytoplasm to the distal end of the growing flagellar structure ...

Cation (Ca²⁺ and Mn²⁺) Partitioning Assays with Intact Arabidopsis Chloroplasts

Authors: Anna Harms, Iris Steinberger and Anja Schneider, date: 01/05/2017, view: 4552, Q&A: 0

[Abstract] Determination of the relative distribution of Ca²⁺ and Mn²⁺ is an important tool for analyzing mutants showing altered levels of calcium and/or manganese transporters in the chloroplast envelope or thylakoid membrane. The method described in this protocol allows quantitative analyses of the relative distribution of calcium ...

Putrescine Biosynthesis Inhibition in Tomato by DFMA and DFMO Treatment

Authors: Emma Fernández-Crespo, Ana Isabel González-Hernández, Loredana Scalschi, Eugenio Llorens, Pilar García-Agustín and Gemma Camañes, date: 11/05/2016, view: 3981, Q&A: 0

[Abstract] This protocol can be used to inhibit the biosynthesis of polyamines, specifically putrescine, in tomato plants grown with NH₄⁺ as a solely N source. In general, polyamines are positively charged small metabolites implicated in physiological processes, including organogenesis, embryogenesis, floral initiation and development, ...

Measurement of Cellular Copper in Rhodobacter capsulatus by Atomic Absorption Spectroscopy

Authors: Petru-Iulian Trasnea, Dorian Marckmann, Marcel Utz and Hans-Georg Koch, date: 10/05/2016, view: 4938, Q&A: 0

[Abstract] Copper is an essential micronutrient and functions as a cofactor in many enzymes such as heme-Cu oxygen reductases, Cu-Zn superoxide dismutases, multi-copper oxidases and tyrosinases. However, due to its chemical reactivity, free copper is highly toxic (Rae et al., 1999) and all organisms use sophisticated machineries for controlling ...

⁵⁹Fe Uptake Assays in Paracoccidioides Species

Authors: Daniel J. Kosman, Elisa Flávia Luiz Cardoso Bailão, Mirelle Garcia Silva-Bailão and Célia Maria de Almeida Soares, date: 09/20/2016, view: 3722, Q&A: 0

[Abstract] Iron is an essential micronutrient required for virtually all organisms. This fact is related to the ability of the transition metal to exist in two oxidation states, the reduced ferrous (Fe²⁺) and the oxidized ferric (Fe³⁺). Given the relative availability of aqueous iron (the element which constitutes ~5% of the earth’s ...

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