JoVE Journal

Medicine

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High-throughput Flow Cytometry Cell-based Assay to Detect Antibodies to N-Methyl-D-aspartate Receptor or Dopamine-2 Receptor in Human Serum

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16.1K Views

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10:19 min

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November 23rd, 2013

DOI :

10.3791/50935-v

November 23rd, 2013

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Mazen Amatoury¹, Vera Merheb¹, Jessica Langer¹, Xin Maggie Wang², Russell Clive Dale¹, Fabienne Brilot¹

¹Institute for Neuroscience and Muscle Research, The Kids Research Institute at the Children's Hospital at Westmead, The University of Sydney, ²Flow Cytometry Centre, Westmead Millennium Institute for Medical Research

Podsumowanie

Over the recent years, live cell-based assays have been used successfully to detect antibodies against surface and conformational antigens. Here, we describe a method using high-throughput flow cytometry enabling the analysis of large cohorts of patients. Detection of novel antibodies will improve diagnosis and treatment of immune-mediated disorders.

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High throughput Flow Cytometry

Cell based Assay

N methyl D aspartate Receptor

Dopamine 2 Receptor

Autoimmune CNS Diseases

Neurotransmission

Eukaryotic Cells

Fluorochrome conjugated Secondary Antibodies

High throughput Sampler System

Antigenic Targets

Central Nervous System

Immune mediated Disorders

Rozdziały w tym wideo

0:05

Title

1:37

Subcloning Strategy: D2R or NMDAR into pIRES2-EGFP Vector

2:20

Transfection: Expression of Neuronal Antigen in HEK293 Cells

4:15

Flow Cytometry Cell-based Assay for Detection of Antibody to Surface Neuronal Antigens

7:44

Results: Detection/Analysis of Antibody Binding

9:45

Conclusion

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