Atanasijevic, Tatjana and Jasanoff, Alan (2007) Preparation of iron oxide-based calcium sensors for MRI. NATURE PROTOCOLS, 2 (10). pp. 2582-2589.
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Official URL: http://www.nature.com/nprot/journal/v2/n10/abs/npr...
We describe a protocol for the preparation of a family of calcium indicators for MRI. The indicators consist of superparamagnetic iron oxide nanoparticles functionalized with the calcium-sensing protein, calmodulin, and its target peptides. Calcium-dependent protein-protein interactions drive particle clustering and produce large changes in T2 relaxivity. To prepare these indicators, interacting polypeptides are purified, biotinylated and then conjugated to streptavidin-coated nanoparticles. Conjugates are isolated, mixed to obtain functional sensors and incubated in the presence or absence of calcium to test response properties. Calcium-dependent responses are assayed by dynamic light scattering and MRI. The whole series of procedures can be carried out in similar to 12 h. The iron oxide-based calcium sensors discussed here are suitable for the detection of calcium concentration changes of similar to 1 mu M in vitro.
|Subjects:||Analytical Science > Nanotechnology for sensing and actuating|
Biomedical Science > Nanobiotechnology
Material Science > Nanochemistry
Material Science > Nanostructured materials
|Deposited By:||Anuj Seth|
|Deposited On:||16 Dec 2008 15:48|
|Last Modified:||19 Jan 2009 12:22|
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