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Spectral evidence for non-calcium interactions of intracellular Indo-1.

Abstract：

:Indo-1 is widely used to measure intracellular free calcium, [Ca2+]i, by comparing the fluorescence emission at 2 or more wavelengths with the emissions, which are assumed to be known, of Indo-1 when it is fully calcium-bound and when it is fully calcium-free. Accurate quantitation requires that these "reference" values be obtained on intracellular dye, and the full spectra of this study show that the reason is a significant spectral shift of the calcium-free peak, but not the calcium-bound. A mathematical analysis shows that the new peak must be a new state of the Indo-1 molecule, since it cannot be simply due to residual calcium in the cell. When intracellular "reference" spectra were used in the data analysis, [Ca2+]i could be calculated from whole spectra or from the ratio of observations at two wavelengths with good agreement. When extracellular "reference" spectra were used, the value calculated by the ratio method depended on the choice of wavelengths.

journal_name

Biochem Biophys Res Commun

journal_title

Biochemical and biophysical research communications

authors

Owen CS,Shuler RL

doi

10.1016/0006-291x(89)92139-6

subject

Has Abstract

pub_date

1989-08-30 00:00:00

pages

328-33

issue

1

eissn

0006-291X

issn

1090-2104

pii

0006-291X(89)92139-6

journal_volume

163

pub_type

杂志文章

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