Fura-2FF-based calcium indicator for protein labeling.
Laboratory of Protein Engineering, Institute of Chemical Sciences and Engineering, Ecole Polytechnique Fédérale de Lausanne (EPFL), CH-1015, Lausanne, Switzerland.Organic & Biomolecular Chemistry (Impact Factor: 3.49). 08/2010; 8(15):3398-401.
ABSTRACT We describe the synthesis and fluorescence properties of a Fura-2FF-based fluorescent Ca(2+) indicator that can be covalently linked to SNAP-tag fusion proteins and retains its Ca(2+) sensing ability after coupling to protein.
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ABSTRACT: Summary form only given. An applied-Br magnetically insulated ion diode is used at Los Alamos for thin film deposition. The authors have measured the radial line-integrated (1 θ 5 cm) electron densities n(z,t) at various z locations within the AK (anode-cathode) gap with two (0.63 and 3.39 μm) heterodyne quadrature laser interferometers. During ion beam emission, no measurable nl (< 1014 cm-2) is observed, except very near K. This suggests that impedance collapses are not caused by A or K plasma closures. At the z location closest (0.3 cm) to K, nl starts to rise 0.1-0.2 μs after beam onset and reaches about 1015 cm-2 by beam fall. These density releases correlate with beam current densities and are delayed with anodes of reduced (2.5 cm) annular thickness. Hence, they appear to be beam-induced K plasmas. After beam emission, large nl's are observed through the entire AK gap. Near K, peak nl's of (2-3) × 1016 cm-2 are measured 1 μs after beam fall. These plasmas appear to propagate from K towards A with average speeds of 0.5 cm/μs. near A, peak nl's reach (6-8) × 1016/ cm-2. These data are consisted with avalanche ionization of flashover-produced neutralsIEEE International Conference on Plasma Science 01/1993;
Article: Subplasma membrane Ca2+ signals.[Show abstract] [Hide abstract]
ABSTRACT: Ca(2+) may selectively activate various processes in part by the cell's ability to localize changes in the concentration of the ion to specific subcellular sites. Interestingly, these Ca(2+) signals begin most often at the plasma membrane space so that understanding subplasma membrane signals is central to an appreciation of local signaling. Several experimental procedures have been developed to study Ca(2+) signals near the plasma membrane, but probably the most prevalent involve the use of fluorescent Ca(2+) indicators and fall into two general approaches. In the first, the Ca(2+) indicators themselves are specifically targeted to the subplasma membrane space to measure Ca(2+) only there. Alternatively, the indicators are allowed to be dispersed throughout the cytoplasm, but the fluorescence emanating from the Ca(2+) signals at the subplasma membrane space is selectively measured using high resolution imaging procedures. Although the targeted indicators offer an immediate appeal because of selectivity and ease of use, their limited dynamic range and slow response to changes in Ca(2+) are a shortcoming. Use of targeted indicators is also largely restricted to cultured cells. High resolution imaging applied with rapidly responding small molecule Ca(2+) indicators can be used in all cells and offers significant improvements in dynamic range and speed of response of the indicator. The approach is technically difficult, however, and realistic calibration of signals is not possible. In this review, a brief overview of local subplasma membrane Ca(2+) signals and methods for their measurement is provided.International Union of Biochemistry and Molecular Biology Life 06/2012; 64(7):573-85. · 2.79 Impact Factor
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ABSTRACT: The development of molecular probes to visualize cellular processes is an important challenge in chemical biology. One possibility to create such cellular indicators is based on the selective labeling of proteins with synthetic probes in living cells. Over the last years, our laboratory has developed different labeling approaches for monitoring protein activity and for localizing synthetic probes inside living cells. In this article, we review two of these labeling approaches, the SNAP-tag and CLIP-tag technologies, and their use for studying cellular processes.CHIMIA International Journal for Chemistry 11/2011; 65(11):868-71. · 1.09 Impact Factor
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