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To perform immunofluorescence staining, a fluorophore must be conjugated (“tagged”) to an antibody. Staining procedures can be applied to both retained intracellular expressed antibodies, or to cell surface antigens on living cells. There are two general classes of immunofluorescence techniques: primary (direct) and secondary (indirect).
Main staining patterns on chromogenic immunohistochemistry. Immunofluorescence of human skin using an anti-IgA antibody. The skin is from a patient with Henoch–Schönlein purpura: IgA deposits are found in the walls of small superficial capillaries (yellow arrows).
Immunohistochemistry or IHC staining of tissue sections (or immunocytochemistry, which is the staining of cells), is perhaps the most commonly applied immunostaining technique. [2] While the first cases of IHC staining used fluorescent dyes (see immunofluorescence ), other non-fluorescent methods using enzymes such as peroxidase (see ...
The location of fluorescence will vary according to the target molecule, external for membrane proteins, and internal for cytoplasmic proteins. In this way immunofluorescence is a powerful technique when combined with confocal microscopy for studying the location of proteins and dynamic processes ( exocytosis , endocytosis , etc.).
Direct FA stained mouse brain impression smear reveals the presence of the bacterium Chlamydia psittaci. 400X.. A direct fluorescent antibody (DFA or dFA), also known as "direct immunofluorescence", [1] is an antibody that has been tagged in a direct fluorescent antibody test.
Immunofluorescence staining pattern of anti-dsDNA antibodies on HEp-20-10 cells (left), Crithidia luciliae (centre) and rat liver (right). Anti-double stranded DNA (Anti-dsDNA) antibodies are a group of anti-nuclear antibodies (ANA) the target antigen of which is double stranded DNA.
Nucleolar staining pattern of ANAs. Until around 1975, when HEp-2 cells were introduced, animal tissue was used as the standard substrate for immunofluorescence. [11] HEp-2 cells are currently one of the most common substrates for ANA detection by immunofluorescence. [54]
Propidium iodide (or PI) is a fluorescent intercalating agent that can be used to stain cells and nucleic acids.PI binds to DNA by intercalating between the bases with little or no sequence preference.
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