New Luminescence-Based Approach to Measurement of Luciferase
Gene Expression Reporter Activity and Adenosine
Triphosphate-Based Determination of Cell Viability

J 2010

New Luminescence-Based Approach to Measurement of Luciferase Gene Expression Reporter Activity and Adenosine Triphosphate-Based Determination of Cell Viability

KONOPKA, R.; M. HYZDALOVA; L. KUBALA a Jiří PACHERNÍK

Základní údaje

Originální název

New Luminescence-Based Approach to Measurement of Luciferase Gene Expression Reporter Activity and Adenosine Triphosphate-Based Determination of Cell Viability

Autoři

KONOPKA, R.; M. HYZDALOVA; L. KUBALA (garant) a Jiří PACHERNÍK (203 Česká republika, domácí)

Vydání

Folia biologica, Prague, Charles University, 2010, 0015-5500

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10608 Biochemistry and molecular biology

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 0.729

Organizační jednotka

Přírodovědecká fakulta

UT WoS

000277599300005

EID Scopus

2-s2.0-77952139966

Klíčová slova anglicky

cell viability; reporter gene assay; luciferase; adenosine triphosphate

Štítky

rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 19. 6. 2020 14:20, Mgr. Marie Novosadová Šípková, DiS.

Anotace

V originále

The assay employing firefly luciferase as the end-point reporter is one of the most popular gene reporter systems. However, the physiological conditions of cells may affect the reporter gene expression, which makes an assessment of cell viability desirable. Estimates of cell viability may be based on different principles. We tested for correlations between various cell viability assessments, including luminescent determination of adenosine triphosphate in whole-cell lysate, and the reporter luciferase activity in pluripotent embryonic and colon adenocarcinoma cells. Luciferase activity in cell lysate from both cell lines cultured under different conditions correlated with the amount of viable cells assessed by all of the methods employed. Importantly, it was also possible to carry out adenosine triphosphate determination in cell lysates prepared in the buffer originally designed for determining luciferase activity; it correlated significantly with adenosine triphosphate determination in cells lysed in the buffer originally designed for adenosine triphosphate determination. The results suggest that the assessment of live cells by determining adenosine triphosphate can be multiplexed with a luciferase reporter gene assay, which allows independent monitoring of both reporter expression and cell viability.

Návaznosti

GA301/08/0717, projekt VaV

Název: Regulace diferenciace ES buněk prostřednictvím receptoru gp130

Investor: Grantová agentura ČR, Regulace diferenciace ES buněk prostřednictvím receptoru gp130

Citovat

KONOPKA, R.; M. HYZDALOVA; L. KUBALA a Jiří PACHERNÍK. New Luminescence-Based Approach to Measurement of Luciferase Gene Expression Reporter Activity and Adenosine Triphosphate-Based Determination of Cell Viability. Folia biologica. Prague: Charles University, 2010, roč. 56, č. 2, s. 66-71. ISSN 0015-5500.

@article{1663182,
   author = {Konopka, R. and Hyzdalova, M. and Kubala, L. and Pacherník, Jiří},
   article_location = {Prague},
   article_number = {2},
   keywords = {cell viability; reporter gene assay; luciferase; adenosine triphosphate},
   language = {eng},
   issn = {0015-5500},
   journal = {Folia biologica},
   title = {New Luminescence-Based Approach to Measurement of Luciferase Gene Expression Reporter Activity and Adenosine Triphosphate-Based Determination of Cell Viability},
   url = {https://fb.cuni.cz/Data/files/folia_biologica/volume_56_2010_2/fb2010A0011.pdf},
   volume = {56},
   year = {2010}
}

TY  - JOUR
ID  - 1663182
AU  - Konopka, R. - Hyzdalova, M. - Kubala, L. - Pacherník, Jiří
PY  - 2010
TI  - New Luminescence-Based Approach to Measurement of Luciferase Gene Expression Reporter Activity and Adenosine Triphosphate-Based Determination of Cell Viability
JF  - Folia biologica
VL  - 56
IS  - 2
SP  - 66-71
EP  - 66-71
PB  - Charles University
SN  - 00155500
KW  - cell viability
KW  - reporter gene assay
KW  - luciferase
KW  - adenosine triphosphate
UR  - https://fb.cuni.cz/Data/files/folia_biologica/volume_56_2010_2/fb2010A0011.pdf
L2  - https://fb.cuni.cz/Data/files/folia_biologica/volume_56_2010_2/fb2010A0011.pdf
N2  - The assay employing firefly luciferase as the end-point reporter is one of the most popular gene reporter systems. However, the physiological conditions of cells may affect the reporter gene expression, which makes an assessment of cell viability desirable. Estimates of cell viability may be based on different principles. We tested for correlations between various cell viability assessments, including luminescent determination of adenosine triphosphate in whole-cell lysate, and the reporter luciferase activity in pluripotent embryonic and colon adenocarcinoma cells. Luciferase activity in cell lysate from both cell lines cultured under different conditions correlated with the amount of viable cells assessed by all of the methods employed. Importantly, it was also possible to carry out adenosine triphosphate determination in cell lysates prepared in the buffer originally designed for determining luciferase activity; it correlated significantly with adenosine triphosphate determination in cells lysed in the buffer originally designed for adenosine triphosphate determination. The results suggest that the assessment of live cells by determining adenosine triphosphate can be multiplexed with a luciferase reporter gene assay, which allows independent monitoring of both reporter expression and cell viability.
ER  -

KONOPKA, R.; M. HYZDALOVA; L. KUBALA a Jiří PACHERNÍK. New Luminescence-Based Approach to Measurement of Luciferase Gene Expression Reporter Activity and Adenosine Triphosphate-Based Determination of Cell Viability. \textit{Folia biologica}. Prague: Charles University, 2010, roč.~56, č.~2, s.~66-71. ISSN~0015-5500.

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