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Preparation of Drosophila Polytene Chromosomes, Followed by Immunofluorescence Analysis of Chromatin Structure by Multi-fluorescence Correlations

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Isolation of Pure Mitochondria from Rat Kidneys and Western Blot of Mitochondrial Respiratory Chain Complexes

Authors: Tamara Micakovic, Wiktoria Z. Banczyk, Euan Clark, Bettina Kränzlin, Jörg Peters and Sigrid Christa Hoffmann, date: 10/05/2019, view: 2048, Q&A: 1

[Abstract] Cardiac, neuronal and renal tubular epithelial cells are the most metabolically active cells in the body. Their fate depends largely on their mitochondria as the primary energy generating system which participates in the control of apoptosis, cell cycle and metabolism. Thus, mitochondrial dysfunction is a hallmark of many chronic diseases ...

Organelle-associated rRNA Degradation

Authors: Jinliang Huang and Geng Wang, date: 06/05/2019, view: 1825, Q&A: 0

[Abstract] Cytosolic rRNAs are highly dynamic and can be degraded under conditions such as apoptosis, starvation and magnesium depletion. The degradation is also related to their specific localization, as fractions of cytosolic ribosomes are localized on the surfaces of intracellular organelles, such as endoplasmic reticulum (ER) and mitochondria. Such ...

Improved Mammalian Mitochondrial RNA Isolation

Authors: Jinliang Huang and Geng Wang, date: 06/05/2019, view: 2602, Q&A: 0

[Abstract] Mitochondria have two sets of RNAs. One is encoded in mitochondrial genome, and the other that consists of imported RNAs within mitochondria and cytosolic RNAs associated with mitochondrial outer membrane is encoded in the nucleus. These mitochondrial RNAs play important roles in mitochondrion biosynthesis and signaling in and out of mitochondria. ...

Detection of Internal Matrix Targeting Signal-like Sequences (iMTS-Ls) in Mitochondrial Precursor Proteins Using the TargetP Prediction Tool

Authors: Felix Boos, Timo Mühlhaus and Johannes M. Herrmann, date: 09/05/2018, view: 3275, Q&A: 0

[Abstract] Mitochondria contain hundreds of proteins which are encoded by the nuclear genome and synthesized in the cytosol from where they are imported into the organelle. Sorting signals encoded in the primary and secondary sequence of these proteins mediate the recognition of newly synthesized precursor proteins and their subsequent translocation through ...

Ca²⁺ Measurements in Mammalian Cells with Aequorin-based Probes

Authors: Anna Tosatto, Rosario Rizzuto and Cristina Mammucari, date: 03/05/2017, view: 5252, Q&A: 0

[Abstract] Aequorin is a Ca²⁺ sensitive photoprotein suitable to measure intracellular Ca²⁺ transients in mammalian cells. Thanks to recombinant cDNAs expression, aequorin can be specifically targeted to various subcellular compartments, thus allowing an accurate measurement of Ca²⁺ uptake and release of different ...

Mouse Liver Mitochondria Isolation, Size Fractionation, and Real-time MOMP Measurement

Authors: Thibaud T. Renault, Mark P.A Luna-Vargas and Jerry E. Chipuk, date: 08/05/2016, view: 7555, Q&A: 0

[Abstract] The mitochondrial pathway of apoptosis involves a complex interplay between dozens of proteins and lipids, and is also dependent on the shape and size of mitochondria. The use of cellular models in past studies has not been ideal for investigating how the complex multi-factor interplay regulates the molecular mechanisms of mitochondrial outer ...

Isolating Liver Mitochondria by Differential Centrifugation

Authors: Ignacio Amigo, Javier Traba and Carlos B. Rueda, date: 05/20/2016, view: 12459, Q&A: 0

[Abstract] In addition to methods aimed at the study of mitochondrial function in-situ, a full understanding of mitochondrial function requires their purification from cells or tissues under specific physiological or pathological conditions. This protocol illustrates a sequential procedure to obtain functional mitochondria with high yield from mice liver ...

Isolating Brain Mitochondria by Differential Centrifugation

Authors: Ignacio Amigo, Javier Traba and Carlos B. Rueda, date: 05/20/2016, view: 11378, Q&A: 3

[Abstract] In addition to methods aimed at the study of mitochondrial function in-situ, a full understanding of mitochondrial function requires their purification from cells or tissues under specific physiological or pathological conditions. This protocol illustrates a sequential procedure to obtain functional mitochondria with high yield from mice ...

Separation of the Inner and Outer Mitochondrial Membrane in HeLa Cells

Authors: Naotaka Nishimura and Masato Yano, date: 11/20/2014, view: 20030, Q&A: 1

[Abstract] Mitochondria are organelles that have important functions in oxidative phosphorylation, fatty acid oxidation and apoptosis signaling. They have two distinct membranes, outer membrane (OM) and inner membrane (IM). IM contains respiratory chain complexes that produce ATP. IM is rich in cardiolipin, a specific phospholipid reportedly having a ...

Isolation of Mitochondria from Potato Tubers

Authors: Jesper F. Havelund, Fernanda Salvato, Mingjie Chen, R.S.P. Rao, Adelina Rogowska-Wrzesinska, Ole N. Jensen, David R. Gang, Jay J. Thelen and Ian Max Møller, date: 09/05/2014, view: 12836, Q&A: 2

[Abstract] One way to study the function of plant mitochondria is to extract them from plant tissues in an uncontaminated, intact and functional form. The reductionist assumption is that the components present in such a preparation and the in vitro measurable functions or activities reliably reflect the in vivo properties of the organelle ...

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