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Extracellular Protein Microarray Technology for High Throughput Detection of Low Affinity Receptor-Ligand Interactions
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Biochemistry
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JoVE Journal Biochemistry
Extracellular Protein Microarray Technology for High Throughput Detection of Low Affinity Receptor-Ligand Interactions

Extracellular Protein Microarray Technology for High Throughput Detection of Low Affinity Receptor-Ligand Interactions

DOI:
10.3791/58451-v

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06:01 min

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January 07, 2019

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, , ,
1Receptor Discovery group, Microchemistry, Proteomics and Lipidomics Department,Genentech, 2Portfolio Management and Operations,Genentech

Chapters

  • 00:04Title
  • 00:28Extracellular Protein Microarray Printing
  • 01:55Preparation of Multivalent Bait Complexes
  • 03:09Extracellular Protein Microarray Screening
  • 04:36Results: Evaluation of Extracellular Protein Microarray Slide Printing and Screen of an Orphan Query Protein
  • 05:11Conclusion

Summary

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Here, we present a protocol to screen extracellular protein microarrays for identification of novel receptor-ligand interactions in high throughput. We also describe a method to enhance detection of transient protein-protein interactions by using protein-microbead complexes.

Tags

Extracellular Protein MicroarrayHigh Throughput DetectionLow Affinity Receptor-ligand InteractionsProtein-protein InteractionsMicro ArrayerEpoxy Silane SlidesCy3 Labeled BSABlocking SolutionMultivalent ApproachFc-tagged Extracellular DomainsProtein A Coated MicrobeadsCy5 IgG Labeling

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