Journal
/
Biochemistry
/
Horizontal Gel Electrophoresis for Enhanced Detection of Protein-RNA Complexes
JoVE Journal
Biochemistry
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biochemistry
Horizontal Gel Electrophoresis for Enhanced Detection of Protein-RNA Complexes

Horizontal Gel Electrophoresis for Enhanced Detection of Protein-RNA Complexes

DOI:
10.3791/56031-v

•

06:36 min

•

July 28, 2017

•

, , , ,
1Department of Biomolecular Chemistry,University of Wisconsin School of Medicine and Public Health, 2Department of Biochemistry and Molecular Pharmacology,University of Massachusetts Medical School

Chapters

  • 00:05Title
  • 00:35Preparing the Horizontal Native Polyacrylamide Gel
  • 02:27Binding Reaction
  • 03:23Analysis of Samples
  • 04:12Imaging the Gel
  • 05:04Results: Horizontal Gel Electrophoresis Enhances the Detection of Distinct RNA-protein Complexes
  • 06:08Conclusion

Summary

Automatic Translation

Automatic Translation

Native polyacrylamide gel electrophoresis is a fundamental tool for analyzing RNA-protein interactions. Traditionally most experiments have used vertical gels. However, horizontal gels provide several advantages, such as the opportunity to monitor complexes during electrophoresis. We provide a detailed protocol for generating and using horizontal native gel electrophoresis.

Tags

Keywords: Horizontal Gel ElectrophoresisProtein-RNA ComplexesRNA BiochemistryRNA-protein InteractionsNative Gel ElectrophoresisGel BoxTBE Running BufferAcrylamide GelBinding ReactionFluorescently Labeled RNABicaudal-c Protein

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/56031?article_type=v

Sheathless Capillary Electrophoresis–Mass Spectrometry for Metabolic Profiling of Biological Samples
check_url/56031?article_type=v

Characterization of Multi-subunit Protein Complexes of Human MxA Using Non-denaturing Polyacrylamide Gel-electrophoresis
check_url/56031?article_type=v

Multimer-PAGE: A Method for Capturing and Resolving Protein Complexes in Biological Samples
check_url/56031?article_type=v

Protein-tRNA Agarose Gel Retardation Assays for the Analysis of the N6-threonylcarbamoyladenosine TcdA Function
check_url/56031?article_type=v

Detection and Visualization of DNA Damage-induced Protein Complexes in Suspension Cell Cultures Using the Proximity Ligation Assay
check_url/56031?article_type=v

Analyzing Supercomplexes of the Mitochondrial Electron Transport Chain with Native Electrophoresis, In-gel Assays, and Electroelution
check_url/56031?article_type=v

DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis
check_url/56031?article_type=v

Use of Two Dimensional Semi-denaturing Detergent Agarose Gel Electrophoresis to Confirm Size Heterogeneity of Amyloid or Amyloid-like Fibers
check_url/56031?article_type=v

Analysis of AtHIRD11 Intrinsic Disorder and Binding Towards Metal Ions by Capillary Gel Electrophoresis and Affinity Capillary Electrophoresis
check_url/56031?article_type=v

Analysis of Thylakoid Membrane Protein Complexes by Blue Native Gel Electrophoresis

Read Article