Mammalia

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    Protocols in Current Issue
    Fluorescence HPLC Analysis of the in-vivo Activity of Glucosylceramide Synthase
    Authors:  Kartik R. Roy, Sachin K. Khiste, Zhijun Liu and Yong-Yu Liu, date: 06/20/2019, view: 1597, Q&A: 0
    [Abstract] Almost all functions of cells or organs rely on the activities of cellular enzymes. Indeed, the in-vivo activities that directly represent the cellular effects of enzymes in live organs are critical importance to appreciate the roles enzymes play in modulating physiological or pathological processes, although assessments of such in-vivo ...
    Cell-free Generation of COPII-coated Procollagen I Carriers
    Authors:  Lin Yuan, Satoshi Baba, Kanika Bajaj and Randy Schekman, date: 11/20/2017, view: 3814, Q&A: 0
    [Abstract] The aim of this protocol is to generate COPII-coated procollagen I (PC1) carriers in a cell-free reaction. The COPII-coated PC1 carriers were reconstituted from donor membrane, cytosol, purified recombinant COPII proteins, and nucleotides. This protocol describes the preparation of donor membrane and cytosol, the assembly of the reaction, and the ...
    Metabolic Heavy Isotope Labeling to Study Glycerophospholipid Homeostasis of Cultured Cells
    Authors:  Satu Hänninen, Pentti Somerharju and Martin Hermansson, date: 05/05/2017, view: 3999, Q&A: 0
    [Abstract] Glycerophospholipids consist of a glycerophosphate backbone to which are esterified two acyl chains and a polar head group. The head group (e.g., choline, ethanolamine, serine or inositol) defines the glycerophospholipid class, while the acyl chains together with the head group define the glycerophospholipid molecular species. Stable ...
    BODIPY 493/503 Staining of Neutral Lipid Droplets for Microscopy and Quantification by Flow Cytometry
    Authors:  Bo Qiu and M. Celeste Simon, date: 09/05/2016, view: 33030, Q&A: 0
    [Abstract] Lipid droplets (LDs) are ubiquitous, dynamic organelles and function as a storage depot for neutral lipids, including triglycerides and cholesterol esters (Walther and Farese, 2012). The movement of lipid species into and out of LDs impacts a variety of cellular processes, such as energy homeostasis, lipid-based signaling, and membrane homeostasis ...
    Determination of Cellular Phosphatidylinositol-3-phosphate (PI3P) Levels Using a Fluorescently Labelled Selective PI3P Binding Domain (PX)
    Authors:  Michael J. Munson and Ian G. Ganley, date: 08/20/2016, view: 4901, Q&A: 0
    [Abstract] The lipid Phosphatidylinositol-3-phosphate [PtdIns3P or PI(3)P] plays many membrane trafficking roles and is primarily produced by the Class III PI3K, VPS34. Determining the level of cellular PI(3)P however can be complex. Extraction of cellular lipids by methanol/chloroform can struggle to separate and identify distinct phospholipid species. ...
    Metabolic Assays for Detection of Neutral Fat Stores
    Authors:  Jan M. Baumann, Leila Kokabee, Xianhiu Wang, Yan Sun, Jason Wong and Douglas S. Conklin, date: 06/20/2015, view: 9628, Q&A: 1
    [Abstract] Lipid droplets (LDs) are ubiquitous intracellular structures whose formation, growth, and maintenance are highly regulated (Wang et al., 2013; Ranall et al., 2011; Goodman, 2009). Lipid metabolism and droplet dynamics are of considerable interest to agriculture, biofuel production, viral pathology, nutrition, and cancer biology ...
    PTEN-lipid Binding Assay
    Authors:  Sridhar Kavela, Swapnil R. Shinde and Subbareddy Maddika, date: 08/20/2013, view: 6961, Q&A: 0
    [Abstract] The lipid and protein interactions are an integral and important part of many cellular signaling pathways. The understanding of the selective and specific interaction of the given lipid molecule with the target protein is required for studying cellular signaling. In this assay, different lipids are spotted onto a nitrocellulose membrane to which ...



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