A live-cell assay for the real-time assessment of extracellular ATP levels

被引:3
作者
Niles, Andrew L. [1 ]
Cali, James J. [1 ]
Lazar, Dan F. [1 ]
机构
[1] Promega Corp, 5430 Cheryl Pkwy, Madison, WI 53711 USA
关键词
Extracellular ATP; eATP; Damage associated molecular patterns; DAMP; Phosphatidylserine PS; Real-time; Non-destructive; Immunogenic cell death; ICD; Apoptosis; Necrosis; FIND-ME SIGNAL; RELEASE; PHOSPHATIDYLSERINE; CHANNELS; DANGER;
D O I
10.1016/j.ab.2021.114286
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Extracellular ATP (eATP) is a potent damage associated molecular pattern (DAMP) molecule known to exert profound effects on the innate and adaptive immune responses. As such, it has become an important biomarker for studying means to pro-actively modulate inflammatory processes. Unfortunately, traditional methodologies employed for measuring eATP require cumbersome supernatant sampling, onerous time courses, or unnecessary duplication of effort. Here we describe a new reagent that is tolerable to test cells in extended exposures and enables a fully homogeneous assay method for real-time determinations of extracellular ATP levels. The reagent is introduced into assay plates containing cells at the time of stimulus introduction. The real-time feature of the format allows for sensitive, continuous accounting of eATP levels in the test model over at least 24 h. This work details our efforts to create and characterize this new reagent and to validate utility by demonstrating its use with multiple cell lines and chemically diverse eATP induction stimuli.
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页数:8
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