Cellular and Subcellular Nanotechnology: Methods and by Daniel F. Moyano, Vincent M. Rotello (auth.), Volkmar

By Daniel F. Moyano, Vincent M. Rotello (auth.), Volkmar Weissig, Tamer Elbayoumi, Mark Olsen (eds.)

In Cellular and Subcellular Nanotechnology: tools and Protocols expert researchers within the box element the newest advances that have been made in using the big power of nanotechnology for probing, imaging and manipulating existence on a mobile and subcellular point. Written within the hugely profitable Methods in Molecular Biology™ sequence layout, chapters contain introductions to their respective subject matters, lists of the mandatory fabrics and reagents, step by step, with ease reproducible laboratory protocols, and key tips about troubleshooting and keeping off identified pitfalls.

Authoritative and functional, Cellular and Subcellular Nanotechnology: equipment and Protocols seeks to assist scientists within the additional examine of utilizing nanotechnology to all parts of biomedical sciences.

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2. Place tubes on a microcentrifuge tube tumbler rotator and rotate at 15 rpm for 12 h at 4°C. 3. Place tubes in the magnetic separator rack for up to 1 h at 4°C. Remove supernatant and resuspend antibody-bead conjugates in sterile PBS. Repeat wash cycle two more times. Finally, resuspend antibody-bead conjugates in 500 mL of sterile PBS and use within 24 h. 2 Treat Cells with Nanoparticles 1. Wash cells three times for 10 min with 37°C serum-free medium to remove residual serum protein. 2. Add nanoparticles in sterile serum-free DMEM (or other appropriate cell medium) to cells at desired concentration (Fig.

Artech House, Norwood MA, pp 433–448 3. Pratusevich VR, Balke CW (1996) Factors shaping the confocal image of the calcium spark in cardiac muscle cells. Biophys J 71: 2942–2957 4. Parfenov AS, Salnikov V, Lederer WJ, Lukyanenko V (2006) Aqueous diffusion pathways as a part of the ventricular cell ultrastructure. Biophys J 90:1107–1119 5. Salnikov VV, Lukyanenko YO, Frederick CA, Lederer WJ, Lukyanenko V (2007) Probing the outer mitochondrial membrane in cardiac mitochondria with nanoparticles. Biophys J 92: 1058–1071 Chapter 5 Immunoisolation of Nanoparticles Containing Endocytic Vesicles for Drug Quantitation Ari Nowacek, Irena Kadiu, JoEllyn McMillan, and Howard E.

2. Cell incubator. 3. Serum-free DMEM (or other appropriate serum-free culture medium). 4. Nanoparticles (see Note 3). 5. Sterile PBS. 3 Homogenization of NanoparticleLoaded Cells 1. 2, 250 mM sucrose, 1 mM EDTA, and 1 mM Mg(OAc)2. 2. Cell scrapers. 3. Dounce homogenizer (7 mL). 4. 15 mL centrifuge tubes. 5. Refrigerated centrifuge. 4 Isolation of NanoparticleContaining Endosomes 1. 3). 2. 1). nanoparticle-treated with attached 3. Magnetic separator rack. 4. Sterile PBS. 5. Refrigerated tabletop centrifuge.

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