Semantic resources project/Antibodies/Revised Methods/Immunofluorescence

Description
IF: Immunofluorescence

Outline
A reaction in which antibodies bind to antigens. The antibodies are themselves attached to a fluorescent dye, or an enzyme which provokes a fluorescent reaction. The result is a color which is visible through microscopy.

Indirect immunofluorescence involves two antibodies: the first specific to the antigen of interest, and the second which binds to the first antibody and carries with it the fluorescent marker.

Usage
Schneider Gasser et al. "Immunofluorescence in brain sections: simultaneous detection of presynaptic and postsynaptic proteins in identified neurons". Nature Protocols 1,1887-1897. (November 2006) "'The classical fixation of brain tissue by intravascular perfusion with an aldehyde solution (or merely by immersion in the fixative) leads to a reduced detection of postsynaptic proteins. By contrast, immunohistochemical staining of vesicle-associated presynaptic proteins, such as vesicular transporters, usually requires excellent tissue preservation — a strong fixation — and therefore the simultaneous detection of presynaptic and postsynaptic markers by immunofluorescence is challenging. Accordingly, there are only a few studies in which satisfactory detection of cell-type markers or neutrotransmitters was combined successfully with immunofluorescence of post-synaptic proteins.'"

Definition
An immunoassay in which the presence of an antigen is detected by means of a fluorescent marker detectable through microscopy. The fluorescent marker is itself attached to the antigen by a bound antibody, or by the sequential binding of two or more antibodies.