5 μL of the recovered blood (not normalized to cell count or recovery, using the cryopreservation/washing procedure described above) was added to 95 μL Alsever’s solution in a 96-well plate. 100 μL CellTiter-Glo 3D Reagent (Promega) was added, and the solution was thoroughly mixed, then incubated at ambient temperatures for 30 min. Luminescence (visible light ~740 to 380 nm) was then measured using a plate reader (Synergy HT, BioTek). Background noise was controlled by subtracting luminescence from samples that were incubated with Alsever’s solution instead of the CellTiter-Glo 3D Reagent. To make the standard curve, 0.011 g adenosine 5’-triphosphate disodium salt hydrate (Thermo Scientific) was added to 1 L of distilled water and diluted via serial dilatation in a 96-well plate. The luciferase assay was conducted as above. The initial 20× dilution of the original samples was accounted for when calculating the final ATP concentration.
Adenosine 5 triphosphate disodium salt hydrate
Manufactured by Thermo Fisher Scientific
Sourced in United States
Adenosine 5'-triphosphate disodium salt hydrate is a chemical compound used in various biochemical and cell-based applications. It is the primary energy-carrying molecule in living organisms, playing a crucial role in cellular processes. This product is commonly used as a reagent in research, drug discovery, and diagnostic workflows.
Automatically generated - may contain errors
Lab products found in correlation
Synergy ht, by Agilent Technologies (2 mentions)
Celltiter glo 3d reagent, by Promega (2 mentions)
2 7 dichlorofluorescin diacetate dcfh da, by Merck Group (1 mentions)
Sodium hydroxide (naoh), by Avantor (1 mentions)
Rpmi 1640 broth medium, by Harvard Bioscience (1 mentions)
Titriplex 3, by Merck Group (1 mentions)
3 protocols using adenosine 5 triphosphate disodium salt hydrate
1
ATP Quantification in Cryopreserved Cells
2
ATP Quantification via Luminescence Assay
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5 μL
of the recovered blood
(not normalized to cell count or recovery, using the cryopreservation/washing
procedure described above) was added to 95 μL Alsever’s
solution in a 96-well plate. 100 μL CellTiter-Glo 3D Reagent
(Promega) was added, and the solution was thoroughly mixed, then incubated
at ambient temperatures for 30 min. Luminescence (visible light ∼740
to 380 nm) was then measured using a plate reader (Synergy HT, BioTek).
Background noise was controlled by subtracting luminescence from samples
that were incubated with Alsever’s solution instead of the
CellTiter-Glo 3D Reagent. To make the standard curve, 0.011 g adenosine
5′-triphosphate disodium salt hydrate (Thermo Scientific) was
added to 1 L of distilled water and diluted via serial dilatation
in a 96-well plate. The luciferase assay was conducted as above. The
initial 20× dilution of the original samples was accounted for
when calculating the final ATP concentration.
of the recovered blood
(not normalized to cell count or recovery, using the cryopreservation/washing
procedure described above) was added to 95 μL Alsever’s
solution in a 96-well plate. 100 μL CellTiter-Glo 3D Reagent
(Promega) was added, and the solution was thoroughly mixed, then incubated
at ambient temperatures for 30 min. Luminescence (visible light ∼740
to 380 nm) was then measured using a plate reader (Synergy HT, BioTek).
Background noise was controlled by subtracting luminescence from samples
that were incubated with Alsever’s solution instead of the
CellTiter-Glo 3D Reagent. To make the standard curve, 0.011 g adenosine
5′-triphosphate disodium salt hydrate (Thermo Scientific) was
added to 1 L of distilled water and diluted via serial dilatation
in a 96-well plate. The luciferase assay was conducted as above. The
initial 20× dilution of the original samples was accounted for
when calculating the final ATP concentration.
3
DMSO-based Cellular Assay Protocol
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Dimethyl sulfoxide (DMSO), 3-(N-morpholino) propane-sulfonic acid (MOPS), resazurin (REZ), tert-butyl hydroperoxide (t-BHP), carbonyl cyanide m-chlorophenyl hydrazone (CCCP), luciferase from Photinus pyralis (firefly) and 2′,7′-dichlorofluorescin diacetate (DCFH-DA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). RPMI-1640 broth medium (with L-glutamine, without bicarbonate and with the pH indicator phenol red) was purchased from Biochrom AG (Berlin, Germany). Phosphate buffered saline (PBS) was purchased from Fisher Reagent (Geel, Belgium). Potassium hydrogen carbonate (KHCO3), sodium dihydrogen phosphate dihydrate (Na2H2PO4.2H2O), adenosine 5′-triphosphate disodium salt hydrate and JC-1 were acquired from Thermo Fisher Scientific (Franklin, MA, USA). Perchloric acid 70% (HClO4) and Titriplex III (EDTA, disodium salt dihydrate) were obtained from Merck (Darmstadt, Germany). Sodium hydroxide (NaOH) was obtained from VWR (Fontenay-sous-Bois, France). D-Luciferin sodium salt was obtained from Abcam (Cambridge, United Kingdom).
Cyclam salt H4[H2(4-CF3PhCH2)2Cyclam]Cl4 (compound1 ) was synthesized according to a previously published procedure [22 (link)].
Cyclam salt H4[H2(4-CF3PhCH2)2Cyclam]Cl4 (compound
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