Gold nanocluster-based fluorescent assay for label-free detection of protein kinase and its inhibitors

Gold nanocluster-based fluorescent assay for label-free detection of protein kinase and its... A sensitive fluorometric assay is reported for label-free detection of the activity of protein kinase (PKA) and of its inhibitors. It is based on the europium(III)-modulated fluorescence of peptide-stabilized gold nanoclusters (peptide-AuNCs). Both adenosine-5′-triphosphate (ATP) and adenosine-5′-diphosphate (ADP) (formed from ATP by PKA-catalyzed hydrolysis) enhance the fluorescence of peptide-AuNCs with its excitation/emission peaks of 330/405 nm. The addition of Eu(III) quenches the fluorescence of the ATP/peptide-AuNC system to the original fluorescence level of peptide-AuNCs, while addition of Eu(III) has no effect on the fluorescence of the ATP/PKA/peptide-AuNC system. Based on this “turn-on” method, the activity of PKA can be detected with high sensitivity in 0.05–1.6 U⋅mL−1 activity range, with a detection limit of 0.02 U⋅mL−1 (3σ). The feasibility of this method for screening of inhibitors was also studied. The IC50 value for the commercially available inhibitor H-89 was found to be 43 nM. Moreover, the detection scheme was also applied to monitoring the drug-stimulated activation of PKA in HepG-2 cell lysates. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Microchimica Acta Springer Journals

Gold nanocluster-based fluorescent assay for label-free detection of protein kinase and its inhibitors

Loading next page...
 
/lp/springer_journal/gold-nanocluster-based-fluorescent-assay-for-label-free-detection-of-lJBQI2nID3
Publisher
Springer Vienna
Copyright
Copyright © 2017 by Springer-Verlag Wien
Subject
Chemistry; Nanochemistry; Nanotechnology; Characterization and Evaluation of Materials; Analytical Chemistry; Microengineering
ISSN
0026-3672
eISSN
1436-5073
D.O.I.
10.1007/s00604-017-2349-2
Publisher site
See Article on Publisher Site

References

You’re reading a free preview. Subscribe to read the entire article.


DeepDyve is your
personal research library

It’s your single place to instantly
discover and read the research
that matters to you.

Enjoy affordable access to
over 12 million articles from more than
10,000 peer-reviewed journals.

All for just $49/month

Explore the DeepDyve Library

Unlimited reading

Read as many articles as you need. Full articles with original layout, charts and figures. Read online, from anywhere.

Stay up to date

Keep up with your field with Personalized Recommendations and Follow Journals to get automatic updates.

Organize your research

It’s easy to organize your research with our built-in tools.

Your journals are on DeepDyve

Read from thousands of the leading scholarly journals from SpringerNature, Elsevier, Wiley-Blackwell, Oxford University Press and more.

All the latest content is available, no embargo periods.

See the journals in your area

Monthly Plan

  • Read unlimited articles
  • Personalized recommendations
  • No expiration
  • Print 20 pages per month
  • 20% off on PDF purchases
  • Organize your research
  • Get updates on your journals and topic searches

$49/month

Start Free Trial

14-day Free Trial

Best Deal — 39% off

Annual Plan

  • All the features of the Professional Plan, but for 39% off!
  • Billed annually
  • No expiration
  • For the normal price of 10 articles elsewhere, you get one full year of unlimited access to articles.

$588

$360/year

billed annually
Start Free Trial

14-day Free Trial