Immunofluorescence (IF) is a powerful method for visualizing proteins expressed directly within tissues. This method combines immunology and fluorescent molecules to identify localized proteins within defined morphological structures and thus, provide insights into gene expression, protein-protein interactions, and biomarker identification. Whether you’re just starting off doing IF or are a seasoned veteran, this guide will take you through the important steps and considerations needed for all immunofluorescence applications.

This guide offers you,

  • An overview of the IF workflow
  • Optimization tips for each step
  • Selection tips for the most appropriate secondary fluorescent conjugates
  • A review of autofluorescence, what it is and how to quench unwanted signal

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Dorsal root ganglia cells (neurons and satelite glia): Beta III tubulin (ms), DyLight® 488 Anti-Mouse IgG, S100 (rb), and DyLight® 594 Anti-Rabbit IgG. Cells mounted in VECTASHIELD® HardSet™ Antifade Mounting Medium with DAPI. Image courtesy of Dr. Emma East, Department of Life Sciences, The Open University, Milton Keynes, UK.