Novel detection method for chemiluminescence derived from the Kinase-Glo luminescent kinase assay platform: Advantages over traditional microplate luminometers

Bell, Robert; Storey, Kenneth B.

doi:10.1016/j.mex.2014.07.003

Cited by 7 publications

(4 citation statements)

References 6 publications

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“…The simplicity of measuring luminescence can be taken advantage of since other types of equipment, such as CCD cameras, can convert light emitted from a luminescent reaction to a digital signal for quantification. Previously it was shown that a Western-blot imaging system can be adapted to reliably detect and quantify luminescent signals [3] . Expanding the use of a common piece of laboratory equipment such as this can be preferable to purchasing a dedicated luminometer, if one is not already present.…”

Section: Methods Detailsmentioning

confidence: 99%

“…If needed, the sensitivity could be increased further by increasing the exposure time, or increasing pixel-binning. We chose to use black microplates to stay consistent with previous methods [3] , however white microplates are often recommended for luminescent assay as they reflect light more efficiently, resulting in greater sensitivity.…”

Section: Additional Informationmentioning

confidence: 99%

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Improved high-throughput quantification of luminescent microplate assays using a common Western-blot imaging system

Hawkins

Storey

2017

MethodsX

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Section: Methods Detailsmentioning

confidence: 99%

Section: Additional Informationmentioning

confidence: 99%

Improved high-throughput quantification of luminescent microplate assays using a common Western-blot imaging system

Hawkins

Storey

2017

MethodsX

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“…It also allows higher ATP concentrations (up to 500 μM), thus making it more versatile. 71 , 93 , 97 , 98 ADP-Quest differs from the two kits described above as it measures fluorescence. ADP-Quest exploits the ADP produced in the kinase reaction to convert 10-acetyl-3,7-dihydroxyphenoxazine (ADHP), a fluorescent dye precursor the fluorescent resorufin.…”

Section: In Vitro Assays For the Evaluation Of Lsrk Activitymentioning

confidence: 99%

“…This assay’s advantages include a more stable luminescence signal and shorter analysis time, making it suitable for the screen of large compound libraries. It also allows higher ATP concentrations (up to 500 μM), thus making it more versatile. ,,, ADP-Quest differs from the two kits described above as it measures fluorescence. ADP-Quest exploits the ADP produced in the kinase reaction to convert 10-acetyl-3,7-dihydroxyphenoxazine (ADHP), a fluorescent dye precursor the fluorescent resorufin.…”

Section: In Vitro Assays For the Evaluation Of Lsrk Activitymentioning

confidence: 99%

Tackling Antimicrobial Resistance with Small Molecules Targeting LsrK: Challenges and Opportunities

et al. 2020

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Antimicrobial resistance (AMR) is a growing threat with severe health and economic consequences. The available antibiotics are losing efficacy, and the hunt for alternative strategies is a priority. Quorum sensing (QS) controls biofilm and virulence factors production. Thus, the quenching of QS to prevent pathogenicity and to increase bacterial susceptibility to antibiotics is an appealing therapeutic strategy. The phosphorylation of autoinducer-2 (a mediator in QS) by LsrK is a crucial step in triggering the QS cascade. Thus, LsrK represents a valuable target in fighting AMR. Few LsrK inhibitors have been reported so far, allowing ample room for further exploration. This perspective aims to provide a comprehensive analysis of the current knowledge about the structural and biological properties of LsrK and the state-of-the-art technology for LsrK inhibitor design. We elaborate on the challenges in developing novel LsrK inhibitors and point out promising avenues for further research.

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Insights into modulating the monastrol scaffold: Development of new pyrimidinones as Eg5 inhibitors with anticancer activity

Thabit

Mostafa

Selim

et al. 2022

Archiv der Pharmazie

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Based on modulation of the monastrol scaffold, two series of pyrimidinone derivatives, 3a-e and 5a-k, were designed, synthesized, and investigated for their in vitro anticancer activity. Compound 5j exhibited the most potent cytotoxic activity against four cancer cell lines, including HCT-116, HeLa, HEPG-2, and MCF-7, with IC 50 values of 3.75-5.13 µM, while proving to be safe in the normal human cell line WI-38, with a selectivity index value of 13.7 on HCT-116 cells. Compounds 3d, 3e, and 5h-j were further assessed for their Eg5 inhibitory activity, where 3d and 5h-j showed high Eg5 inhibition with IC 50 values of 28.48, 24.22, 18.90, and 12.89 µM, respectively, when compared to monastrol (IC 50 = 14.89 µM). Cell cycle distribution of HCT-116 cells monitored with compound 5j illustrated that the cell cycle was arrested at the G2/M phase, with considerable apoptotic effect.A molecular docking study was performed to investigate the mode of action of the synthesized anticancer agents as Eg5 inhibitors.

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Novel detection method for chemiluminescence derived from the Kinase-Glo luminescent kinase assay platform: Advantages over traditional microplate luminometers

Abstract: Graphical abstract

Cited by 7 publications

References 6 publications

Improved high-throughput quantification of luminescent microplate assays using a common Western-blot imaging system

Improved high-throughput quantification of luminescent microplate assays using a common Western-blot imaging system

Tackling Antimicrobial Resistance with Small Molecules Targeting LsrK: Challenges and Opportunities

Insights into modulating the monastrol scaffold: Development of new pyrimidinones as Eg5 inhibitors with anticancer activity

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