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Microwaving and Fluorophore-Tyramide for Multiplex Immunostaining on Mouse Adrenals − Using Unconjugated Primary Antibodies from the Same Host Species
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Biologia dello sviluppo
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JoVE Journal Biologia dello sviluppo
Microwaving and Fluorophore-Tyramide for Multiplex Immunostaining on Mouse Adrenals − Using Unconjugated Primary Antibodies from the Same Host Species

Microwaving and Fluorophore-Tyramide for Multiplex Immunostaining on Mouse Adrenals − Using Unconjugated Primary Antibodies from the Same Host Species

DOI:
10.3791/60868-v

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07:37 min

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February 21, 2020

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, , ,
1College of Animal Science & Technology,Henan University of Science and Technology, 2Department of Anatomy, Physiology and Pharmacology, College of Veterinary Medicine,Auburn University

Capitoli

  • 00:05Introduction
  • 00:42First Target Protein Staining
  • 03:15Antibody Stripping
  • 03:41Second Target Protein Staining
  • 04:49Fluorescence Microscopy Imaging
  • 05:30Results: Representative Effects of Microwave-Mediated Stripping on Antibody Cross-Reactivity
  • 06:38Conclusion

Summary

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Several different methods have been established for multiplex immunostaining using primary antibodies from the same host species. Here, we describe the use of microwave-mediated antibody stripping and of fluorophore-tyramide to block antibody cross-reactivity during multiplex immunostaining on formalin-fixed paraffin-embedded mouse adrenal sections.

Tags

Multiplex ImmunostainingUnconjugated Primary AntibodiesFormalin-fixed Paraffin-embeddedSodium CitratePBSTBlocking SolutionSecondary AntibodyStreptavidin Horseradish PeroxidaseFluorophore-tyramide

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