Human Peripheral Blood Neutrophil Isolation for Interrogating the Parkinson's Associated LRRK2 Kinase Pathway by Assessing Rab10 Phosphorylation

7.3K Views

•

12:49 min

•

March 21st, 2020

DOI :

10.3791/58956-v

March 21st, 2020

•

Ying Fan*¹, Francesca Tonelli*¹, Shalini Padmanabhan², Marco A.S. Baptista², Lindsey Riley², Danielle Smith³, Connie Marras⁴, Andrew Howden⁵, Dario R. Alessi¹, Esther Sammler¹^,⁶

¹MRC Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of Dundee, ²The Michael J. Fox Foundation for Parkinson's Research, ³Department of Medical and Molecular Genetics, Indiana University School of Medicine, ⁴Morton and Gloria Shulman Movement Disorders Centre and the Edmond J. Safra Program in Parkinson's Disease, Toronto Western Hospital, University of Toronto, ⁵Division of Cell Signalling and Immunology, School of Life Sciences, University of Dundee, ⁶Division of Molecular and Clinical Medicine, School of Medicine, Ninewells Hospital and Medical School, University of Dundee

Summary

Explore More Videos

Neutrophil Isolation

LRRK2 Kinase

Rab10 Phosphorylation

Endogenous LRRK2 Activity

Human Peripheral Blood

Phospho specific Antibodies

Chapters in this video

0:01

Introduction

0:48

Neutrophil Isolation from Human Peripheral Blood

7:18

LRRK2 Kinase Inhibitor Treatment

10:33

Results

12:14

Conclusion

Related Videos

08:05

Assaying Protein Kinase Activity with Radiolabeled ATP

18.2K Views

09:16

Oligopeptide Competition Assay for Phosphorylation Site Determination

8.3K Views

08:55

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

14.8K Views

09:22

Protein Digestion, Ultrafiltration, and Size Exclusion Chromatography to Optimize the Isolation of Exosomes from Human Blood Plasma and Serum

16.4K Views

12:26

Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay

18.4K Views

10:05

Studying RNA Interactors of Protein Kinase RNA-Activated during the Mammalian Cell Cycle

6.4K Views

08:00

Enhancing the Engraftment of Human Induced Pluripotent Stem Cell-derived Cardiomyocytes via a Transient Inhibition of Rho Kinase Activity

6.1K Views

08:53

Translating Ribosome Affinity Purification (TRAP) for RNA Isolation from Endothelial Cells In Vivo

11.7K Views

13:22

Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays

7.8K Views

06:49

Nonradioactive Assay to Measure Polynucleotide Phosphorylation of Small Nucleotide Substrates

4.6K Views