Transport —Cell-based analysis —Cell Biology

An in vitro Blood-brain Barrier Model to Study the Penetration of Nanoparticles

Authors: Sezin Aday, Wen Li, Jeffrey M. Karp and Nitin Joshi, date: 02/20/2022, view: 2558, Q&A: 0

[Abstract]

The blood-brain barrier (BBB), a crucial protection mechanism in the central nervous system (CNS), is a selective barrier comprised of endothelial cells. It hampers the development of therapeutic and diagnostic tools for neurological diseases due to the poor penetration of most of these agents. Rationally engineered nanoparticles (NP) can

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Polyamine Transport Assay Using Reconstituted Yeast Membranes

Authors: Sarah van Veen, Shaun Martin, Marleen Schuermans and Peter Vangheluwe, date: 01/20/2021, view: 2239, Q&A: 0

[Abstract]

ATP13A2/PARK9 is a late endo-/lysosomal P5B transport ATPase that is associated with several neurodegenerative disorders. We recently characterized ATP13A2 as a lysosomal polyamine exporter, which sheds light on the molecular identity of the unknown mammalian polyamine transport system. Here, we describe step by step a protocol to measure

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K⁺ Release Assay and K⁺ Measurement in Oocyte Assay

Author: Hong Li, date: 11/05/2020, view: 2051, Q&A: 0

[Abstract] The Xenopus oocyte is a powerful system for the exogenous expression and functional characterization of plant membrane transport proteins. Until now, a number of potassium transporters and channels have been identified in oocytes expression system by the two-electrode voltage clamp technology. It is difficult to characterize K⁺ ...

Stopped-flow Light Scattering Analysis of Red Blood Cell Glycerol Permeability

Authors: Patrizia Gena, Piero Portincasa, Sabino Matera, Yonathan Sonntag, Michael Rutzler and Giuseppe Calamita, date: 08/20/2020, view: 2640, Q&A: 0

[Abstract] Stopped-Flow Light Scattering (SFLS) is a method devised to analyze the kinetics of fast chemical reactions that result in a significant change of the average molecular weight and/or in the shape of the reaction substrates. Several modifications of the original stopped-flow system have been made leading to a significant extension of its technical ...

Electroporation of Labeled Antibodies to Visualize Endogenous Proteins and Posttranslational Modifications in Living Metazoan Cell Types

Authors: Sascha Conic, Dominique Desplancq, László Tora and Etienne Weiss, date: 11/05/2018, view: 4202, Q&A: 0

[Abstract] The spatiotemporal localization of different intracellular factors in real-time and their detection in live cells are important parameters to understand dynamic protein-based processes. Therefore, there is a demand to perform live-cell imaging and to measure endogenous protein dynamics in single cells. However, fluorescent labeling of endogenous ...

Transcytosis Assay for Transport of Glycosphingolipids across MDCK-II Cells

Authors: Maria Daniela Garcia-Castillo, Wayne I. Lencer and Daniel J.-F. Chinnapen, date: 10/20/2018, view: 5225, Q&A: 0

[Abstract] Absorption and secretion of peptide and protein cargoes across single-cell thick mucosal and endothelial barriers occurs by active endocytic and vesicular trafficking that connects one side of the epithelial or endothelial cell (the lumen) with the other (the serosa or blood). Assays that assess this pathway must robustly control for non-specific ...

Boron Uptake Assay in Xenopus laevis Oocytes

Authors: Sheliang Wang, Namiki Mitani-Ueno and Junpei Takano, date: 03/05/2018, view: 6459, Q&A: 0

[Abstract] Boron (B) is essential for plant growth and taken up by plant roots as boric acid. Under B limitation, B uptake and translocation in plants are dependent on the boric acid channels located in the plasma membrane. Xenopus leavis oocyte is a reliable heterologous expression system to characterize transport activities of boric acid channels ...

Microvesicle Isolation from Rat Brain Extract Treated Human Mesenchymal Stem Cells

Authors: Ji Yong Lee, Seong-Mi Choi and Han-Soo Kim, date: 07/05/2017, view: 9430, Q&A: 0

[Abstract] Microvesicle (MVs) are submicron-sized membranous vesicles that are either actively released from cells via secretory compartments or shed from cell surface membranes. MVs are generated by many cell types and serve as vehicles that transfer biological information (e.g., protein, mRNA, and miRNA) to distant cells, thereby affecting their ...

Analysis of Mitochondrial Transfer in Direct Co-cultures of Human Monocyte-derived Macrophages (MDM) and Mesenchymal Stem Cells (MSC)

Authors: Megan V. Jackson and Anna D. Krasnodembskaya, date: 05/05/2017, view: 11027, Q&A: 0

[Abstract] Mesenchymal stem/stromal cells (MSC) are adult stem cells which have been shown to improve survival, enhance bacterial clearance and alleviate inflammation in pre-clinical models of acute respiratory distress syndrome (ARDS) and sepsis. These diseases are characterised by uncontrolled inflammation often underpinned by bacterial infection. The ...

In vitro Assays for Measuring Endothelial Permeability by Transwells and Electrical Impedance Systems

Authors: Hong-Ru Chen and Trai-Ming Yeh, date: 05/05/2017, view: 23033, Q&A: 2

[Abstract] Vascular leakage is an important feature in several diseases, such as septic shock, viral hemorrhagic fever, cancer metastasis and ischemia-reperfusion injuries. Thus establishing assays for measuring endothelial permeability will provide insight into the establishment or progression of such diseases. Here, we provide transwell permeability assay ...

Cell Biology

Protocol for Isolation of Cardiomyocyte from Adult Mouse and Rat

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