Immunofluorescence technology is also known as fluorescent antibody technology. According to the principle of antigen antibody reaction, the known antigen or antibody is labeled with fluorescent group, and then the fluorescent antibody (or antigen) is used as a probe to detect the corresponding antigen (or antibody) in cells or tissues. The cells or tissues where the fluorescence is located can be seen by fluorescence microscope, so as to determine the nature and location of antigen or antibody.
Endocrine hormones, proteins, polypeptides, nucleic acids, neurotransmitters, receptors, cytokines, cell surface antigens, tumor markers, blood concentration and other bioactive substances were determined.
1.Cell preparation
2.Fixed.According to the need to select the appropriate fixative to fix the cells, the fixed cells could be stored in PBS containing azide for 3 months at 4 ℃
3.Transparent.Cells fixed with crosslinking agents (such as paraformaldehyde) usually need to be incubated with antibodies.In order to ensure that the antibody can reach the antigen site, the cells were treated with permeability before.The nature of antigen protein should be considered in the selection of permeabilizer.
4.Close.The cells were sealed with blocking solution.
5.First antibody binding.
6.Secondary antibody binding.Indirect immunofluorescence requires the use of secondary antibodies.
7.Cover.Fluorescence microscopy.
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