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Related Concept Videos

Laser Microdissection-Based Protocol for the LC-MS/MS Analysis of the Proteomic Profile of Neuromelanin Granules07:35

Laser Microdissection-Based Protocol for the LC-MS/MS Analysis of the Proteomic Profile of Neuromelanin Granules

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A robust protocol is presented here for isolating neuromelanin granules from human post-mortem substantia nigra pars compacta tissue via laser microdissection. This revised and optimized protocol massively minimizes the required time for sample collection, reduces the required sample amount, and enhances the identification and quantification of proteins by LC-MS/MS...
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Hydrolysis of ATP01:08

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The bonds of adenosine triphosphate (ATP) can be broken through the addition of water, releasing one or two phosphate groups in an exergonic process called hydrolysis. This reaction liberates the energy in the bonds for use in the cell—for instance, to synthesize proteins from amino acids.
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Acyl-RAC (Acyl-Resin Assisted Capture) is a highly sensitive, reliable and easy to perform method to detect reversible lipid modification of cysteine residues (S-acylation) in a variety of biological...
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The present protocol provides detailed descriptions for the efficient isolation of urinary extracellular vesicles utilizing functionalized magnetic beads. Moreover, it encompasses subsequent analyses, including western blotting, proteomics, and...
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ATP Yield01:31

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Cellular respiration produces 30 - 32 ATP per glucose molecule. Although most of the ATP results from oxidative phosphorylation and the electron transport chain (ETC), 4 ATP are gained beforehand (2 from glycolysis and 2 from the citric acid cycle).
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Generation of Microtumors Using 3D Human Biogel Culture System and Patient-derived Glioblastoma Cells for Kinomic Profiling and Drug Response Testing09:24

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Patient-derived xenografts of glioblastoma multiforme can be miniaturized into living microtumors using 3D human biogel culture system. This in vivo-like 3D tumor assay is suitable for drug response testing and molecular profiling, including kinomic analysis.
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Related Experiment Video

Updated: Jan 20, 2026

Laser Microdissection-Based Protocol for the LC-MS/MS Analysis of the Proteomic Profile of Neuromelanin Granules
07:35

Laser Microdissection-Based Protocol for the LC-MS/MS Analysis of the Proteomic Profile of Neuromelanin Granules

Published on: December 16, 2021

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Activity-Based Kinome Profiling Using Chemical Proteomics and ATP Acyl Phosphates.

Caroline E Franks1, Ku-Lung Hsu1

  • 1Department of Chemistry, University of Virginia, Charlottesville, Virginia.

Current Protocols in Chemical Biology
|September 5, 2019
PubMed
Summary

This study introduces activity-based probes to assess the potency and selectivity of kinase inhibitors, crucial for developing safer drugs targeting human kinases involved in disease.

Keywords:
KiNativactivity-based protein profilingchemical proteomicschemoproteomicskinasekinomemass spectrometry

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Last Updated: Jan 20, 2026

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Author Spotlight: Advancing EVtrap for High-Throughput Proteomics in Disease Biomarker Discovery
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Area of Science:

  • Biochemistry
  • Pharmacology
  • Proteomics

Background:

  • Human kinases (500+) regulate cell biology via ATP-dependent phosphorylation.
  • Kinase dysregulation and mutations are linked to human diseases.
  • Assessing kinase inhibitor selectivity is vital to prevent drug toxicity due to off-target effects.

Purpose of the Study:

  • To outline protocols for evaluating kinase inhibitor potency and selectivity.
  • To introduce ATP acyl phosphate activity-based probes for chemical proteomic assays.

Main Methods:

  • Utilizing fluorescent gel-based detection methods.
  • Employing mass spectrometry-based detection methods.
  • Implementing competitive chemical proteomic assays with activity-based probes.

Main Results:

  • Developed protocols to evaluate kinase inhibitor potency and selectivity.
  • Demonstrated the ability to assess engagement of >200 native kinase targets.
  • Enabled direct evaluation within complex proteomes.

Conclusions:

  • ATP acyl phosphate activity-based probes provide a robust method for kinase inhibitor profiling.
  • These assays are critical for assessing the selectivity of drugs targeting the human kinome.
  • The described methods facilitate the development of safer and more effective kinase-targeted therapies.