Journal
/
Biochemistry
/
Rab10 Phosphorylation Detection by LRRK2 Activity Using SDS-PAGE with a Phosphate-binding Tag
JoVE Journal
Biochemistry
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biochemistry
Rab10 Phosphorylation Detection by LRRK2 Activity Using SDS-PAGE with a Phosphate-binding Tag

Rab10 Phosphorylation Detection by LRRK2 Activity Using SDS-PAGE with a Phosphate-binding Tag

DOI:
10.3791/56688-v

•

08:55 min

•

December 14, 2017

•

,
1Laboratory of Brain and Neurological Disorders, Graduate School of Pharmaceutical Sciences,The University of Tokyo, 2Laboratory of Neuropathology and Neuroscience, Graduate School of Pharmaceutical Sciences,The University of Tokyo

Chapters

  • 00:05Title
  • 00:53Collecting Cell Lysates
  • 02:34SDS-PAGE and Gel Transfer
  • 04:41Immunoblotting
  • 06:22Results: P-tag Gel Analysis
  • 07:50Conclusion

Summary

Automatic Translation

Automatic Translation

The present study describes a simple method of detecting endogenous levels of Rab10 phosphorylation by leucine-rich repeat kinase 2.

Tags

Keywords: Rab10 PhosphorylationLRRK2 ActivitySDS-PAGEPhosphate-binding TagParkinson’s ResearchHEK293 CellsCell LysisBradford AssaySample PreparationSDS-PAGE GelManganese Chloride

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/56688?article_type=v

Highly Sensitive and Rapid Fluorescence Detection with a Portable FRET Analyzer
check_url/56688?article_type=v

Detection of the pH-dependent Activity of Escherichia coli Chaperone HdeB In Vitro and In Vivo
check_url/56688?article_type=v

Assaying Protein Kinase Activity with Radiolabeled ATP
check_url/56688?article_type=v

Oligopeptide Competition Assay for Phosphorylation Site Determination
check_url/56688?article_type=v

DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis
check_url/56688?article_type=v

Detection of Small GTPase Prenylation and GTP Binding Using Membrane Fractionation and GTPase-linked Immunosorbent Assay
check_url/56688?article_type=v

Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay
check_url/56688?article_type=v

Affinity Purification of Chloroplast Translocon Protein Complexes Using the TAP Tag
check_url/56688?article_type=v

Human Peripheral Blood Neutrophil Isolation for Interrogating the Parkinson’s Associated LRRK2 Kinase Pathway by Assessing Rab10 Phosphorylation
check_url/56688?article_type=v

Detection of Protein Ubiquitination Sites by Peptide Enrichment and Mass Spectrometry

Read Article