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

Authors :
Inouye S
Hojo H
Source :
Biochemical and biophysical research communications [Biochem Biophys Res Commun] 2018 Dec 09; Vol. 507 (1-4), pp. 242-245. Date of Electronic Publication: 2018 Nov 10.
Publication Year :
2018

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 I <subscript>max</subscript> was 7.5-fold higher than that of reAequorin and was calculated to be 3.8 × 10 <superscript>16</superscript> quanta/mg protein.<br /> (Copyright © 2018 Elsevier Inc. All rights reserved.)

Details

Language :
English
ISSN :
1090-2104
Volume :
507
Issue :
1-4
Database :
MEDLINE
Journal :
Biochemical and biophysical research communications
Publication Type :
Academic Journal
Accession number :
30424878
Full Text :
https://doi.org/10.1016/j.bbrc.2018.11.015