Calcium Signaling Protocols by Alec W. M. Simpson (auth.), David G. Lambert (eds.)

By Alec W. M. Simpson (auth.), David G. Lambert (eds.)

Now greatly up-to-date and elevated, Calcium Signaling Protocols, moment variation keeps the distinction of the extremely popular first version. New to this version are chapters at the electrophysiological size of Ca2+ channel task, greater assurance of confocal microscopy, and functional tutorials on of the commonest commercial high-throughput machines, the Fluorometric Imaging Plate Reader (FLIPR), and the FlexStation. All experimental chapters supply with no trouble reproducible strategies for measuring the various elements of Ca2+ signaling. The protocols stick to the profitable tools in Molecular Biology™ sequence layout, each one providing step by step laboratory directions, an advent outlining the rules at the back of the method, lists of the required gear and reagents, and tips about troubleshooting and fending off recognized pitfalls.
Authoritative and up to date, Calcium Signaling Protocols, moment version presents for amateur and professional researchers alike either theoretical and sensible info for the optimum layout and function of reviews of intracellular Ca2+ regulation.

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Chem. 275, 25,892–25,899. 77. Green, A. , and Simpson A. W. M. (2002) Atrial natriuretic peptide attenuates Ca2+ oscillations in rat hepatocytes by modulating plasma membrane Ca2+ fluxes. Gastroenterol. 123, 1291–1203. 78. M. , and Pozzan, T. (1992) Rapid changes of mitochondrial Ca2+ revealed by specifically targeted recombinant aequorin. Nature 358, 325–327. 79. , and Pozzan, T. (2000) Recombinant aequorin as a tool for monitoring calcium concentration in subcellular compartments. Methods Enzymol.

Natl. Acad. Sci. USA 98, 3197–3202. 98. , and Imoto, K. (2001) A high signal-to-noise Ca2+ probe composed of a single green fluorescent protein. Nat. Biotechnol. 19, 137–141. 99. , et al. (2001) Beat-to-beat oscillations of mitochondrial [Ca2+] in cardiac cells EMBO J. 20, 4998–5007. 100. Schild, D. (1996) Laser scanning microscopy and calcium imaging. Cell Calcium 19, 281–296. 101. Parker, I. and Marriott, G. ) (2003) Biophotonics, Part A. in Methods in Enzymology, vol. 360, Academic Press, San Diego.

Here, the indicator is excited at a longer wavelength and either two or three coincident photons (depending on the dye and excitation wavelength) are able to excite the indicator. Indo-1 is normally excited at approx 350 nm, but can also be excited by light close to 700 nm. The resolution over conventional imaging is enhanced, because statistically the arrival of coincident photons only occurs in a very narrow focal plane. Excitation by longer wavelengths reduces autofluorescence and photobleaching, and therefore, the technique has some advantages over other methods.

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