Journal
/
Biochemistry
/
Nonradioactive Assay to Measure Polynucleotide Phosphorylation of Small Nucleotide Substrates
JoVE Journal
Biochemistry
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biochemistry
Nonradioactive Assay to Measure Polynucleotide Phosphorylation of Small Nucleotide Substrates

Nonradioactive Assay to Measure Polynucleotide Phosphorylation of Small Nucleotide Substrates

DOI:
10.3791/61258-v

•

06:49 min

•

May 08, 2020

•

,
1Signal Transduction Laboratory, National Institute of Environmental Health Sciences, Department of Health and Human Services,National Institutes of Health

Chapters

  • 00:05Introduction
  • 00:29In Vitro RNA Kinase Reaction
  • 01:27Gel Electrophoresis
  • 04:53Results: Quantification of RNA Phosphorylation
  • 06:06Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol describes a nonradioactive assay to measure kinase activity of polynucleotide kinases (PNKs) on small DNA and RNA substrates.

Tags

Keywords: Polynucleotide KinasePhosphorylationDNARNASubstrateNonradioactive AssayAcrylamide GelDenaturing GelUreaTBE BufferRNA Enzyme Kinase ReactionsATP SubstrateQuenchingDownstream Analysis

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/61258?article_type=v

Oligopeptide Competition Assay for Phosphorylation Site Determination
check_url/61258?article_type=v

Quantitative Immunofluorescence to Measure Global Localized Translation
check_url/61258?article_type=v

Assay for Phosphorylation and Microtubule Binding Along with Localization of Tau Protein in Colorectal Cancer Cells
check_url/61258?article_type=v

Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay
check_url/61258?article_type=v

Proofreading and DNA Repair Assay Using Single Nucleotide Extension and MALDI-TOF Mass Spectrometry Analysis
check_url/61258?article_type=v

Dual DNA Rulers to Study the Mechanism of Ribosome Translocation with Single-Nucleotide Resolution
check_url/61258?article_type=v

Use of Microscale Thermophoresis to Measure Protein-Lipid Interactions
check_url/61258?article_type=v

Using an Extracellular Flux Analyzer to Measure Changes in Glycolysis and Oxidative Phosphorylation during Mouse Sperm Capacitation
check_url/61258?article_type=v

Measuring Nucleotide Binding to Intact, Functional Membrane Proteins in Real Time
check_url/61258?article_type=v

A High-Throughput Enzyme-Coupled Activity Assay to Probe Small Molecule Interaction with the dNTPase SAMHD1

Read Article