Cell Biology

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    Protocols in Current Issue
    Image-based Quantification of Macropinocytosis Using Dextran Uptake into Cultured Cells
    Authors:  Anh H. Le and Laura M. Machesky, date: 04/05/2022, view: 1069, Q&A: 0
    [Abstract]

    Macropinocytosis is an evolutionarily conserved process, which is characterized by the formation of membrane ruffles and the uptake of extracellular fluid. We recently demonstrated a role for CYFIP-related Rac1 Interactor (CYRI) proteins in macropinocytosis. High-molecular weight dextran (70kDa or higher) has generally been used as a marker for

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    Measurement of Reactive Oxygen and Nitrogen Species in Living Cells Using the Probe 2',7'-Dichlorodihydrofluorescein
    Authors:  Waleska Dornas and Detlef Schuppan, date: 12/20/2021, view: 1487, Q&A: 0
    [Abstract]

    Reactive oxygen species and reactive nitrogen species (RONS) are involved in programmed cell death in the context of numerous degenerative and chronic diseases. In particular, the ability of cells to maintain redox homeostasis is necessary for an adaptive cellular response to adverse conditions that can cause damage to proteins and DNA, resulting

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    Immunoprecipitation of Acetyl-lysine and Western Blotting of Long-chain acyl-CoA Dehydrogenases and Beta-hydroxyacyl-CoA Dehydrogenase in Palmitic Acid Treated Human Renal Tubular Epithelial Cells
    Authors:  Tingting Lv, Suwei Zhu, Yuan Ma, Hong Feng and Qiang Wan, date: 09/20/2020, view: 3264, Q&A: 0
    [Abstract] As one of the main energy metabolism organs, kidney has been proved to have high energy requirements and are more inclined to fatty acid metabolism as the main energy source. Long-chain acyl-CoA dehydrogenases (LCAD) and beta-hydroxyacyl-CoA dehydrogenase (beta-HAD), key enzymes involved in fatty acid oxidation, has been identified as the ...
    Gas Chromatography Detection Protocol of Short-chain Fatty Acids in Mice Feces
    Authors:  Chaozheng Zhang, An Liu, Tianshuang Zhang, Yi Li and Hua Zhao, date: 07/05/2020, view: 3797, Q&A: 0
    [Abstract] Short-chain fatty acids (SCFAs), which are formed mainly by bacteria fermenting undigested carbohydrates in the colon, they are based on the number of carbon atoms in the carbon chain. Organic fatty acids with less than 6 carbon atoms are called short-chain fatty acids. SCFAs are closely related to various aspects of the human body, so more and ...
    Spectrophotometric Assessment of Heme Oxygenase-1 Activity in Leishmania-infected Macrophages
    Authors:  Moumita Basu, Shriya Saha and Anindita Ukil, date: 04/05/2020, view: 3593, 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: 4543, 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
    [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
    [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
    [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: 64775, Q&A: 1
    [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 ...



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