Ionic effects on firefly bioluminescence assay of red blood cell ATP

Louis M. Aledort; Robert I. Weed; Stanley B. Troup

doi:10.1016/0003-2697(66)90205-3

Ionic effects on firefly bioluminescence assay of red blood cell ATP

Louis M. Aledort
, Robert I. Weed
, Stanley B. Troup

Research output: Contribution to journal › Article › peer-review

47 Scopus citations

Abstract

1. (1) Light emission mediated by firefly luciferin-luciferase and ATP is inhibited by increasing the ionic strength of the medium. 2. (2) The inhibition of firefly luciferase by ions has been found to vary with the ion used. Cation inhibition occurs in the following order: Ca⁺⁺ > K⁺ > Na⁺ > Rb⁺ > Li⁺ > choline⁺. Anion inhibition occurs in the following order: I^- > H₂PO₄^- > Br^- > ClO₃^- > Fl^- > Fl^- > HCO₃^- > COOCH₃·H₂O^-. 3. (3) Inhibition of light emission by Na⁺ and K⁺ at 110 mM ionic strength is characterized by maximal emission at concentrations of 87 mM Na⁺ and 23 mM K⁺. 4. (4) 10^-4M ouabain does not affect light emission. 5. (5) Use of the luciferase assay for RBC ATP requires careful control of the ionic composition of the assay. Adequate extraction of ATP requires either boiling or TCA extraction with a volume ratio of 20 1, TCA RBC.

Original language	English
Pages (from-to)	268-277
Number of pages	10
Journal	Analytical Biochemistry
Volume	17
Issue number	2
DOIs	https://doi.org/10.1016/0003-2697(66)90205-3
State	Published - Nov 1966
Externally published	Yes

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@article{2dec5f0e657a4b1cbbf66ef4c030f87f,

title = "Ionic effects on firefly bioluminescence assay of red blood cell ATP",

abstract = "1. (1) Light emission mediated by firefly luciferin-luciferase and ATP is inhibited by increasing the ionic strength of the medium. 2. (2) The inhibition of firefly luciferase by ions has been found to vary with the ion used. Cation inhibition occurs in the following order: Ca++ > K+ > Na+ > Rb+ > Li+ > choline+. Anion inhibition occurs in the following order: I- > H2PO4- > Br- > ClO3- > Fl- > Fl- > HCO3- > COOCH3·H2O-. 3. (3) Inhibition of light emission by Na+ and K+ at 110 mM ionic strength is characterized by maximal emission at concentrations of 87 mM Na+ and 23 mM K+. 4. (4) 10-4M ouabain does not affect light emission. 5. (5) Use of the luciferase assay for RBC ATP requires careful control of the ionic composition of the assay. Adequate extraction of ATP requires either boiling or TCA extraction with a volume ratio of 20 1, TCA RBC.",

author = "Aledort, \{Louis M.\} and Weed, \{Robert I.\} and Troup, \{Stanley B.\}",

note = "Funding Information: The authors would like to thank Dr. Claude Reecl who carried c\&actions and adenine and phosphorus determinations for us. This work was supported by the United States Public Health number l-FZ-HE-28-604-01, Grants HE-O-6241-05, HE-04167-07, United States Atomic Energy Commission at University Energy Project, Rochester, Xew Pork.",

year = "1966",

month = nov,

doi = "10.1016/0003-2697(66)90205-3",

language = "English",

volume = "17",

pages = "268--277",

journal = "Analytical Biochemistry",

issn = "0003-2697",

publisher = "Academic Press Inc.",

number = "2",

}

TY - JOUR

T1 - Ionic effects on firefly bioluminescence assay of red blood cell ATP

AU - Aledort, Louis M.

AU - Weed, Robert I.

AU - Troup, Stanley B.

N1 - Funding Information: The authors would like to thank Dr. Claude Reecl who carried c&actions and adenine and phosphorus determinations for us. This work was supported by the United States Public Health number l-FZ-HE-28-604-01, Grants HE-O-6241-05, HE-04167-07, United States Atomic Energy Commission at University Energy Project, Rochester, Xew Pork.

PY - 1966/11

Y1 - 1966/11

N2 - 1. (1) Light emission mediated by firefly luciferin-luciferase and ATP is inhibited by increasing the ionic strength of the medium. 2. (2) The inhibition of firefly luciferase by ions has been found to vary with the ion used. Cation inhibition occurs in the following order: Ca++ > K+ > Na+ > Rb+ > Li+ > choline+. Anion inhibition occurs in the following order: I- > H2PO4- > Br- > ClO3- > Fl- > Fl- > HCO3- > COOCH3·H2O-. 3. (3) Inhibition of light emission by Na+ and K+ at 110 mM ionic strength is characterized by maximal emission at concentrations of 87 mM Na+ and 23 mM K+. 4. (4) 10-4M ouabain does not affect light emission. 5. (5) Use of the luciferase assay for RBC ATP requires careful control of the ionic composition of the assay. Adequate extraction of ATP requires either boiling or TCA extraction with a volume ratio of 20 1, TCA RBC.

AB - 1. (1) Light emission mediated by firefly luciferin-luciferase and ATP is inhibited by increasing the ionic strength of the medium. 2. (2) The inhibition of firefly luciferase by ions has been found to vary with the ion used. Cation inhibition occurs in the following order: Ca++ > K+ > Na+ > Rb+ > Li+ > choline+. Anion inhibition occurs in the following order: I- > H2PO4- > Br- > ClO3- > Fl- > Fl- > HCO3- > COOCH3·H2O-. 3. (3) Inhibition of light emission by Na+ and K+ at 110 mM ionic strength is characterized by maximal emission at concentrations of 87 mM Na+ and 23 mM K+. 4. (4) 10-4M ouabain does not affect light emission. 5. (5) Use of the luciferase assay for RBC ATP requires careful control of the ionic composition of the assay. Adequate extraction of ATP requires either boiling or TCA extraction with a volume ratio of 20 1, TCA RBC.

UR - https://www.scopus.com/pages/publications/0013970008

U2 - 10.1016/0003-2697(66)90205-3

DO - 10.1016/0003-2697(66)90205-3

M3 - Article

C2 - 5971421

AN - SCOPUS:0013970008

SN - 0003-2697

VL - 17

SP - 268

EP - 277

JO - Analytical Biochemistry

JF - Analytical Biochemistry

IS - 2

ER -

Ionic effects on firefly bioluminescence assay of red blood cell ATP

Abstract

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