Journal
/
Biology
/
Metabolic Labeling of Leucine Rich Repeat Kinases 1 and 2 with Radioactive Phosphate
JoVE Journal
Biology
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biology
Metabolic Labeling of Leucine Rich Repeat Kinases 1 and 2 with Radioactive Phosphate

Metabolic Labeling of Leucine Rich Repeat Kinases 1 and 2 with Radioactive Phosphate

DOI:
10.3791/50523-v

•

11:31 min

•

September 18, 2013

•

, ,
1Laboratory for Neurobiology and Gene Therapy, Department of Neurosciences,KU Leuven and Leuven Institute for Neuroscience and Disease (LIND)

Chapters

  • 00:05Title
  • 02:02Metabolic Labeling of Cells
  • 05:50Analysis of Protein Labeling
  • 09:46Results: Metabolic Labeling of LRRK1 and LRRK2
  • 10:57Conclusion

Summary

Automatic Translation

Automatic Translation

Leucine rich repeat kinases 1 and 2 (LRRK1 and LRRK2) are multidomain proteins which encode both GTPase and kinase domains and which are phosphorylated in cells. Here, we present a protocol to label LRRK1 and LRRK2 in cells with 32P orthophosphate, thereby providing a means to measure their overall cellular phophorylation levels.

Tags

Keywords: Metabolic LabelingLeucine Rich Repeat Kinases 1 And 2 (LRRK1LRRK2)Radioactive PhosphateProtein PhosphorylationImmunoprecipitationSDS-PAGEAutoradiographyWestern Blotting

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/50523?article_type=v

Labeling Stem Cells with Fluorescent Dyes for non-invasive Detection with Optical Imaging
check_url/50523?article_type=v

Studying Membrane Biogenesis with a Luciferase-Based Reporter Gene Assay
check_url/50523?article_type=v

Lineage Labeling of Zebrafish Cells with Laser Uncagable Fluorescein Dextran
check_url/50523?article_type=v

Imaging Glycans in Zebrafish Embryos by Metabolic Labeling and Bioorthogonal Click Chemistry
check_url/50523?article_type=v

Metabolic Pathway Confirmation and Discovery Through 13C-labeling of Proteinogenic Amino Acids
check_url/50523?article_type=v

Measuring Cell Cycle Progression Kinetics with Metabolic Labeling and Flow Cytometry
check_url/50523?article_type=v

A Protocol for the Identification of Protein-protein Interactions Based on 15N Metabolic Labeling, Immunoprecipitation, Quantitative Mass Spectrometry and Affinity Modulation
check_url/50523?article_type=v

Metabolic Labeling of Newly Transcribed RNA for High Resolution Gene Expression Profiling of RNA Synthesis, Processing and Decay in Cell Culture
check_url/50523?article_type=v

Metabolic Labeling and Membrane Fractionation for Comparative Proteomic Analysis of Arabidopsis thaliana Suspension Cell Cultures
check_url/50523?article_type=v

A New Approach for the Comparative Analysis of Multiprotein Complexes Based on 15N Metabolic Labeling and Quantitative Mass Spectrometry

Read Article