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Related Concept Videos

Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
Photoluminescence: Applications01:14

Photoluminescence: Applications

Photoluminescence offers a wide range of applications due to its inherent sensitivity and selectivity. This technique allows for both direct and indirect analyses of the analyte. Direct quantitative analysis is possible when the analyte exhibits a favorable quantum yield for fluorescence or phosphorescence. However, an indirect analysis may be feasible if the analyte is not fluorescent or phosphorescent, or if the quantum yield is unfavorable. Indirect methods include reacting the analyte with...
Labeling DNA Probes03:31

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
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Related Experiment Video

Updated: Jul 6, 2026

Assaying Protein Kinase Activity with Radiolabeled ATP
08:05

Assaying Protein Kinase Activity with Radiolabeled ATP

Published on: May 26, 2017

Fluorescence detection techniques for protein kinase assay.

YongJin Li1, WeiHong Xie, GuiJie Fang

  • 1College of Bioengineering, Hubei University of Technology, Wuhan, 430068, China. yongjin.li@yahoo.com.cn

Analytical and Bioanalytical Chemistry
|March 15, 2008
PubMed
Summary

Non-radioactive fluorescence detection techniques offer a safer and more stable alternative to traditional radioactive methods for protein kinase (PK) assays. This review explores four fluorescence methods for PK assay development.

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Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein
11:23

Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein

Published on: June 30, 2019

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Last Updated: Jul 6, 2026

Assaying Protein Kinase Activity with Radiolabeled ATP
08:05

Assaying Protein Kinase Activity with Radiolabeled ATP

Published on: May 26, 2017

Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein
11:23

Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein

Published on: June 30, 2019

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Molecular Biology

Background:

  • Traditional protein kinase (PK) assays rely on radioactive (32)P-labeled adenosine triphosphate (ATP).
  • Radioactive methods present challenges including hazardous waste generation and the short half-life of (32)P-labeled ATP.
  • Non-radioactive techniques are sought as alternatives for PK assay development.

Purpose of the Study:

  • To review the principles of four fluorescence detection techniques applicable to PK assays.
  • To provide an overview of the applications of these fluorescence techniques in PK assays.
  • To highlight the advantages, limitations, and future research trends in fluorescence-based PK assays.

Main Methods:

  • Fluorescence intensity endpoint measurement
  • Fluorescence resonance energy transfer (FRET)
  • Fluorescence polarization (FP)
  • Fluorescence lifetime imaging

Main Results:

  • Fluorescence techniques provide viable non-radioactive alternatives for PK assays.
  • Each fluorescence method possesses distinct advantages and limitations for PK detection.
  • Research trends indicate a shift towards advanced fluorescence-based PK assay methodologies.

Conclusions:

  • Fluorescence detection techniques are effective alternatives to radioactive methods for protein kinase assays.
  • Understanding the principles and applications of these techniques is crucial for assay optimization.
  • Continued research in fluorescence-based assays promises further advancements in PK research.