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Revalidation of recombinant aequorin as a light emission standard: Estimation of specific activity of Gaussia luciferase

Author:
Inouye, Satoshi, Hojo, Hironobu
Source:
Biochemical and biophysical research communications 2018 v.507 no.1-4 pp. 242-245
ISSN:
0006-291X
Subject:
absorbance, aequorin, amino acid composition, detection limit, light intensity, luciferase, luminescence
Abstract:
To validate the use of recombinant aequorin (reAequorin) as a light emission standard, the protein concentrations of highly purified reAequorin were determined by amino acid composition analysis, and the presence of active reAequorin was confirmed by the ratio of absorbance peak at 460 nm to that at 280 nm. The high correlation of the luminescence intensity with the protein concentration showed that reAequorin could be used for a light emission standard to study the luminescence properties of luciferases and to evaluate the detection sensitivity of luminometers. The specific activity of Gaussia luciferase with Imax was 7.5-fold higher than that of reAequorin and was calculated to be 3.8 × 1016 quanta/mg protein.
Agid:
6225467