Staining antibodies protocol

Immunostaining with Fluorescent Antibodies

Uses for immunofluorescence (IF)—where an antibody is conjugated to a molecule that fluoresces when excited by lasers— include protein localization, confirmation of post-translational modification or activation, and proximity to/complexing with other proteins. Immunocytochemistry (ICC) is a well-established technique that uses antibodies to bind to targets in cell samples, and Coons et al first described immunodetection using a fluorescent reporter molecule in 1942.  In addition to providing information about subcellular targets, combining immunofluorescence with immunocytochemistry produces some of the most compelling visual data in the life sciences.   In this guide, antibody scientists share what we’ve learned about getting the best possible image from your IF-ICC experiments