INDIRECT IMMUNOFLUORESCENCE Meaning and
Definition
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Indirect immunofluorescence is a laboratory technique used to visualize specific proteins or antigens in a sample, such as cells or tissues, through the use of fluorescently-labeled antibodies. It is a valuable tool in immunology, molecular biology, and medical diagnostics.
The process begins with the preparation of a sample, which could be a cultured cell line, tissue section, or patient specimen. The sample is first incubated with a primary antibody, which specifically binds to the desired protein or antigen of interest. This primary antibody is not directly labeled with a fluorescent dye.
After washing away any unbound primary antibody, a secondary antibody is introduced. The secondary antibody is typically raised against the species in which the primary antibody was produced, such as rabbit anti-mouse or goat anti-rabbit. Importantly, the secondary antibody is conjugated with a fluorophore, a molecule that emits fluorescence when excited by light of a specific wavelength.
Upon binding to the primary antibody, the secondary antibody brings the fluorophore into close proximity to the target protein or antigen. When the sample is illuminated with a light of the corresponding wavelength, the fluorophore absorbs the light energy and emits a fluorescent glow, indicating the presence and location of the protein or antigen.
Indirect immunofluorescence allows for the amplification of the signal, as multiple secondary antibodies can bind to a single primary antibody, resulting in a stronger fluorescence signal. This technique enables researchers to observe and study the distribution, localization, and abundance of specific proteins or antigens within cells or tissues.
Common Misspellings for INDIRECT IMMUNOFLUORESCENCE
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- indurect immunofluorescence
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Etymology of INDIRECT IMMUNOFLUORESCENCE
The word "indirect immunofluorescence" has its etymology rooted in the combination of several terms:
1. Indirect: The term "indirect" in this context refers to the indirect method used in immunofluorescence. It implies that an intermediate step or reagent is used to detect the presence of a specific antibody or antigen. This indirect method involves the use of a secondary antibody that is labeled with a fluorescent dye and binds to the primary antibody or antigen of interest.
2. Immunofluorescence: "Immunofluorescence" combines two words - "immuno" and "fluorescence". "Immuno" refers to the immune system or immunity, while "fluorescence" relates to the emission of light by a substance when exposed to certain wavelengths.
