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Immunofluorescence (IF) is a common morphological approach used to determine the distribution of subcellular components. Antibodies that conjugated with fluorescent dyes are required in IF assay. The antibody specifically recognizes the antigen by binding to the epitope of target, and the fluorophore will be detected under a fluorescent microscope.

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immunofluorescence guide

Immunofluorescence Guide

Immunofluorescence (IF) is a common morphological approach used to determine

the distribution of subcellular components. Antibodies that conjugated with

fluorescent dyes are required in IF assay. The antibody specifically recognizes the

antigen by binding to the epitope of target, and the fluorophore will be detected

under a fluorescent microscope. Hence, subcellular components can be visualized in

a dark background. IF can also be used as an alternative semiquantitative analysis

method to monitoring the expression of the interest.

There are three types of IF: direct IF, indirect IF and combined IF.

Direct IF is using a single primary antibody that is conjugated with fluorescent dye.

Indirect IF is using two antibodies for the staining: primary antibody that specifically

binds to epitope and a matched secondary antibody conjugated with fluorescence

dye.

Combined IF is a combination of direct and indirect IF staining.

https://www.creative-diagnostics.com/immunofluorescence-guide.htm