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

You might also read

Related Articles

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

Sort by
Same author

Evolutionary-based remodeling of ABA receptors reveals the structural basis of hormone perception and regulation.

Proceedings of the National Academy of Sciences of the United States of America·2026
Same author

From wing movements to cues and signals: mechanisms and functions of flight-generated sounds in insects.

The Journal of experimental biology·2026
Same author

Integrated multi-omics analyses reveal that p-coumaroyl-CoA 2'-hydroxylases act upstream of stilbene synthases to mediate oxyresveratrol biosynthesis in mulberry (Morus alba).

Plant communications·2026
Same author

Bewitching the host: Striga disrupts host stomatal ABA signaling by impairing the tryptophan lock.

Journal of experimental botany·2026
Same author

Abscisic Acid Enhances Ex Vitro Acclimatization Performance in Hop (<i>Humulus lupulus</i> L.).

International journal of molecular sciences·2025
Same author

PME10 Is a Pectin Methylesterase Driving PME Activity and Immunity Against Botrytis cinerea in Grapevine (Vitis vinifera L.).

Plant biotechnology journal·2025
Same journal

Editorial: Technologies for RNA Detection.

Bio-protocol·2026
Same journal

One-Step Affinity Purification of MarathonRT Reverse Transcriptase for RNA Sequencing Applications.

Bio-protocol·2026
Same journal

Enhanced RNA-Seq Expression Profiling and Functional Enrichment in Non-model Organisms Using Custom Annotations.

Bio-protocol·2026
Same journal

Using Combined Fluorescent In Situ Hybridization With Immunohistochemistry to Co-localize mRNA in Diverse Neuronal Cell Types.

Bio-protocol·2026
Same journal

Stepwise Protocol for Alternative Splicing Analysis in Single-Cell SMART-Seq2 RNA-Seq Data.

Bio-protocol·2026
Same journal

Enriching Bacteria-Specific RNA From Host Samples Before NGS With Transcript-Capture.

Bio-protocol·2026
See all related articles

Related Experiment Video

Updated: Oct 22, 2025

Assaying Protein Kinase Activity with Radiolabeled ATP
08:05

Assaying Protein Kinase Activity with Radiolabeled ATP

Published on: May 26, 2017

18.7K

In Gel Kinase Assay.

Gastón A Pizzio1, Pedro L Rodriguez2

  • 1Department of Molecular Genetics, Centre for Research in Agricultural Genomics (CRAG; consortium CSIC-IRTA-UAB-UB), Barcelona, Spain.

Bio-Protocol
|August 30, 2021
PubMed
Summary
This summary is machine-generated.

A new In Gel Kinase Assay protocol measures protein kinase activity. This assay revealed that the Arabidopsis sextuple PYR/PYL/RCAR-ABA receptor mutant impairs abscisic acid (ABA)-mediated activation of SnRK2 kinases.

Keywords:
ABAArabidopsisEnzymatic activityKinasesPhosphorylationSnRKsin vivo

More Related Videos

Identification of Kinase-substrate Pairs Using High Throughput Screening
11:13

Identification of Kinase-substrate Pairs Using High Throughput Screening

Published on: August 29, 2015

8.4K
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

6.4K

Related Experiment Videos

Last Updated: Oct 22, 2025

Assaying Protein Kinase Activity with Radiolabeled ATP
08:05

Assaying Protein Kinase Activity with Radiolabeled ATP

Published on: May 26, 2017

18.7K
Identification of Kinase-substrate Pairs Using High Throughput Screening
11:13

Identification of Kinase-substrate Pairs Using High Throughput Screening

Published on: August 29, 2015

8.4K
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

6.4K

Area of Science:

  • Plant molecular biology
  • Cellular signaling
  • Biochemistry

Background:

  • Proper spatiotemporal regulation of protein phosphorylation is crucial for cellular development and homeostasis.
  • Abscisic acid (ABA) signaling pathways are vital for plant responses to environmental stress.
  • SnRK2 kinases play a key role in ABA signaling.

Purpose of the Study:

  • To present a novel 'In Gel Kinase Assay' protocol for measuring protein kinase activity in crude extracts.
  • To investigate the role of PYRABACTIN RESISTANCE1/PYR1-LIKE/REGULATORY COMPONENTS OF ABA RECEPTORS (PYR/PYL/RCAR-ABA) in ABA-mediated SnRK2 activation in Arabidopsis thaliana.

Main Methods:

  • Development and application of the 'In Gel Kinase Assay' protocol.
  • Analysis of protein kinase activity in crude protein extracts from wild-type and mutant Arabidopsis lines.
  • Comparison of ABA-mediated activation of SnRK2.2, SnRK2.3, and OST1/SnRK2.6 in sextuple and triple mutants.

Main Results:

  • The 'In Gel Kinase Assay' protocol effectively measures kinase activity from crude protein extracts.
  • The Arabidopsis sextuple mutant (pyr/pyl112458) exhibits impaired ABA-mediated activation of SnRK2.2, SnRK2.3, and OST1/SnRK2.6.
  • This impairment in the sextuple mutant is comparable to that observed in the triple mutant (snrk2.2/2.3/2.6).

Conclusions:

  • The 'In Gel Kinase Assay' is a valuable tool for assessing kinase activity.
  • PYR/PYL/RCAR-ABA receptors are essential for the proper ABA-mediated activation of SnRK2 kinases in Arabidopsis.
  • This finding deepens the understanding of ABA signaling pathways in plants.