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Optimized bioluminescence analysis of adenosine triphosphate (ATP) released by platelets and its application in the high throughput screening of platelet inhibitors

Activated platelets release adenosine trisphosphate (ATP) and bioluminescence analysis of ATP release is usually used to monitor activation of platelets induced by various stimulants. However, bioluminescence analysis of ATP possesses poor linearity, the signal is quickly attenuated, and the accurac...

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Autores principales: Wang, Lili, Li, Yunqian, Guo, Ran, Li, Shanshan, Chang, Anqi, Zhu, Zhixiang, Tu, Pengfei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6786574/
https://www.ncbi.nlm.nih.gov/pubmed/31600247
http://dx.doi.org/10.1371/journal.pone.0223096
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author Wang, Lili
Li, Yunqian
Guo, Ran
Li, Shanshan
Chang, Anqi
Zhu, Zhixiang
Tu, Pengfei
author_facet Wang, Lili
Li, Yunqian
Guo, Ran
Li, Shanshan
Chang, Anqi
Zhu, Zhixiang
Tu, Pengfei
author_sort Wang, Lili
collection PubMed
description Activated platelets release adenosine trisphosphate (ATP) and bioluminescence analysis of ATP release is usually used to monitor activation of platelets induced by various stimulants. However, bioluminescence analysis of ATP possesses poor linearity, the signal is quickly attenuated, and the accuracy of ATP release from platelets is hard to determine accurately enough to be used in a high throughput screening of platelet inhibitors. The present study was designed to optimize bioluminescence analysis of ATP released by platelets and expand its application in high throughput screening of platelet inhibitors. The results showed that accuracy of ATP analysis was significantly improved by adding coenzyme A (CoA) and signal attenuation of ATP analysis was greatly postponed by adding bovine serum albumin (BSA) both in Hank’s balanced salt solution (HBSS) and Tyrode’s buffer. Furthermore, ATP release of activated platelets and inhibitory effects of Ly294002 and Staurosporine on platelet activation were accurately determined by our optimized bioluminescence analysis of ATP. Thus, we have successfully constructed an optimized bioluminescence analysis of ATP which can be used in high throughput screening of platelet inhibitors.
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spelling pubmed-67865742019-10-19 Optimized bioluminescence analysis of adenosine triphosphate (ATP) released by platelets and its application in the high throughput screening of platelet inhibitors Wang, Lili Li, Yunqian Guo, Ran Li, Shanshan Chang, Anqi Zhu, Zhixiang Tu, Pengfei PLoS One Research Article Activated platelets release adenosine trisphosphate (ATP) and bioluminescence analysis of ATP release is usually used to monitor activation of platelets induced by various stimulants. However, bioluminescence analysis of ATP possesses poor linearity, the signal is quickly attenuated, and the accuracy of ATP release from platelets is hard to determine accurately enough to be used in a high throughput screening of platelet inhibitors. The present study was designed to optimize bioluminescence analysis of ATP released by platelets and expand its application in high throughput screening of platelet inhibitors. The results showed that accuracy of ATP analysis was significantly improved by adding coenzyme A (CoA) and signal attenuation of ATP analysis was greatly postponed by adding bovine serum albumin (BSA) both in Hank’s balanced salt solution (HBSS) and Tyrode’s buffer. Furthermore, ATP release of activated platelets and inhibitory effects of Ly294002 and Staurosporine on platelet activation were accurately determined by our optimized bioluminescence analysis of ATP. Thus, we have successfully constructed an optimized bioluminescence analysis of ATP which can be used in high throughput screening of platelet inhibitors. Public Library of Science 2019-10-10 /pmc/articles/PMC6786574/ /pubmed/31600247 http://dx.doi.org/10.1371/journal.pone.0223096 Text en © 2019 Wang et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Wang, Lili
Li, Yunqian
Guo, Ran
Li, Shanshan
Chang, Anqi
Zhu, Zhixiang
Tu, Pengfei
Optimized bioluminescence analysis of adenosine triphosphate (ATP) released by platelets and its application in the high throughput screening of platelet inhibitors
title Optimized bioluminescence analysis of adenosine triphosphate (ATP) released by platelets and its application in the high throughput screening of platelet inhibitors
title_full Optimized bioluminescence analysis of adenosine triphosphate (ATP) released by platelets and its application in the high throughput screening of platelet inhibitors
title_fullStr Optimized bioluminescence analysis of adenosine triphosphate (ATP) released by platelets and its application in the high throughput screening of platelet inhibitors
title_full_unstemmed Optimized bioluminescence analysis of adenosine triphosphate (ATP) released by platelets and its application in the high throughput screening of platelet inhibitors
title_short Optimized bioluminescence analysis of adenosine triphosphate (ATP) released by platelets and its application in the high throughput screening of platelet inhibitors
title_sort optimized bioluminescence analysis of adenosine triphosphate (atp) released by platelets and its application in the high throughput screening of platelet inhibitors
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6786574/
https://www.ncbi.nlm.nih.gov/pubmed/31600247
http://dx.doi.org/10.1371/journal.pone.0223096
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