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Colorectal Cancer Cell Surface Protein Profiling Using an Antibody Microarray and Fluorescence Multiplexing
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Immunologia e infezioni
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JoVE Journal Immunologia e infezioni
Colorectal Cancer Cell Surface Protein Profiling Using an Antibody Microarray and Fluorescence Multiplexing

Colorectal Cancer Cell Surface Protein Profiling Using an Antibody Microarray and Fluorescence Multiplexing

DOI:
10.3791/3322-v

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15:17 min

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September 25, 2011

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, , , , ,
1School of Molecular Bioscience,University of Sydney, 2Department of Surgery,Royal Prince Alfred Hospital, 3Department of Anatomical Pathology,Department of Anatomical Pathology, 4Department of Medicine,Concord Repatriation General Hospital

Capitoli

  • 00:05Titolo
  • 02:00Introduction
  • 03:38Sample Digestion
  • 04:39Sample Filtration
  • 06:32Sample Preparation for Cell Capture
  • 08:18Antibody Microarray Cell Capture
  • 10:41Fluorescence Multiplexing with Conjugated Antibodies
  • 12:38Risultati
  • 14:37Conclusion

Summary

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We described a procedure for the disaggregation of colorectal cancer (CRC) to produce viable single cells, which are then captured on customized antibody microarrays recognizing surface antigens (DotScan CRC microarray). Sub-populations of cells bound to the microarray can be profiled by fluorescence multiplexing using monoclonal antibodies tagged with fluorescent dyes.

Tags

Colorectal CancerCell Surface Protein ProfilingAntibody MicroarrayFluorescence MultiplexingCRC PrognosisProtein ExpressionPost-translational ModificationCluster Of Differentiation AntigensBiomarkersImmunohistochemistry (IHC)Cancer Sub-classificationPrognosticationMarkerClinico-pathological StagingProteomic TechniquesITRAQDiagnostic Laboratories

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