Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

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...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Functional regulation of the protein phosphatase PPM1M by phosphorylation at multiple sites with Ser/Thr-Pro motifs.

Archives of biochemistry and biophysics·2024
Same author

Subcellular distribution of bone morphogenetic protein 2-inducible kinase (BMP2K): Regulation by liquid-liquid phase separation and nucleocytoplasmic shuttling.

Biochemical and biophysical research communications·2023
Same author

CaMK phosphatase (CaMKP/POPX2/PPM1F) inhibitors suppress the migration of human breast cancer MDA-MB-231 cells with loss of polarized morphology.

Biochemical and biophysical research communications·2022
Same author

CaM kinase phosphatase (CaMKP/PPM1F/POPX2) is specifically inactivated through gallate-mediated protein carbonylation.

Archives of biochemistry and biophysics·2022
Same author

Biochemical characterization of four splice variants of mouse Ca2+/calmodulin-dependent protein kinase Iδ.

Journal of biochemistry·2021
Same author

Dual phosphorylation of protein phosphatase PPM1H promotes dephosphorylation of Smad1 in cellulo.

Biochemical and biophysical research communications·2020

Related Experiment Video

Updated: May 30, 2026

Nonradioactive Assay to Measure Polynucleotide Phosphorylation of Small Nucleotide Substrates
06:49

Nonradioactive Assay to Measure Polynucleotide Phosphorylation of Small Nucleotide Substrates

Published on: May 8, 2020

In-gel phosphatase assay using fluorogenic and radioactive substrates.

Isamu Kameshita1

  • 1Kagawa University, Kagawa, Japan.

Current Protocols in Protein Science
|August 16, 2011
PubMed
Summary

This study details two in-gel phosphatase assays for analyzing cellular signaling. These methods, using fluorogenic or radiolabeled substrates, help investigate protein phosphorylation regulation by detecting phosphatase activity.

More Related Videos

A Purification and In Vitro Activity Assay for a (p)ppGpp Synthetase from Clostridium difficile
09:53

A Purification and In Vitro Activity Assay for a (p)ppGpp Synthetase from Clostridium difficile

Published on: November 3, 2018

Use of Recombinant Fusion Proteins in a Fluorescent Protease Assay Platform and Their In-gel Renaturation
19:23

Use of Recombinant Fusion Proteins in a Fluorescent Protease Assay Platform and Their In-gel Renaturation

Published on: January 16, 2019

Related Experiment Videos

Last Updated: May 30, 2026

Nonradioactive Assay to Measure Polynucleotide Phosphorylation of Small Nucleotide Substrates
06:49

Nonradioactive Assay to Measure Polynucleotide Phosphorylation of Small Nucleotide Substrates

Published on: May 8, 2020

A Purification and In Vitro Activity Assay for a (p)ppGpp Synthetase from Clostridium difficile
09:53

A Purification and In Vitro Activity Assay for a (p)ppGpp Synthetase from Clostridium difficile

Published on: November 3, 2018

Use of Recombinant Fusion Proteins in a Fluorescent Protease Assay Platform and Their In-gel Renaturation
19:23

Use of Recombinant Fusion Proteins in a Fluorescent Protease Assay Platform and Their In-gel Renaturation

Published on: January 16, 2019

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cellular Signaling

Background:

  • Protein phosphorylation is a key regulatory mechanism in cellular signaling.
  • Understanding protein phosphatases and kinases is crucial for studying these pathways.

Purpose of the Study:

  • To describe and compare two distinct in-gel phosphatase assay methods.
  • To provide tools for analyzing phosphatase activity in cellular and tissue samples.

Main Methods:

  • In-gel phosphatase assay using fluorogenic substrates (e.g., MUP, DiFMUP) after native-PAGE.
  • In-gel phosphatase assay using (32)P-labeled substrates after SDS-PAGE, followed by renaturation and autoradiography.

Main Results:

  • Both assays allow for in situ detection of phosphatase activities.
  • Each method offers specific advantages and disadvantages for analyzing dephosphorylation.

Conclusions:

  • These in-gel assays are valuable for investigating the roles of protein phosphatases in cellular regulation.
  • The choice of assay depends on the specific research question and sample type.