Subcellular localization immunofluorescence

Immunofluorescence To identify protein-expressing cells in the population and subcellular localization of express protein Relatively efficient and provides intraceUnlar details A qualitative method requiring cell fixation, which inactivates the protein expressing cells... 

Pollenz RS, Sattler CA, Poland A. 1994. The aryl hydrocarbon receptor and aryl hydrocarbon receptor nuclear translocator protein show distinct subcellular localizations in Hepa lclc7 cells by immunofluorescence microscopy. Mol Pharmacol 45 428-438. 

A fluorescent antibody study of the calcium binding protein location has suggested that it is formed in the goblet cells and is secreted from the goblet cells along the surface of the columnar epithelial tissues A wider dispersion of this protein has been suggested from other immunofluorescent antibody studies but the question of specificity of antibody must be taken into account. The exact cellular and subcellular localization of calcium binding protein is as of this date unresolved. 

We have used isolated nucleoli to prepare monoclonal antibodies against nucleolar proteins (S. Chen, J. E. Dove, and J. P. Aris, unpublished results). Six monoclonal antibodies have been characterized by indirect immunofluorescence localization and Western blotting, and have been used to screen yeast expression libraries. At this stage, putative, novel nucleolar proteins are being expressed in yeast with an epitope tag for the purpose of confirming their subcellular localization. 

Immunofluorescence staining permits detection of protein antigens in situ, in order to investigate the subcellular localization or cellular distribution within a tissue. The cells or tissue sections are fixed and incubated with the specific primary monoclonal antibody. The antigen-primaiy monoclonal antibody complex is bound by a second antibody conjugated to a fluorescent dye, such as rhodamine-p-isothiocyanate or fluorescein isothiocyanate, for detection by fluorescence microscopy. Immunofluorescence staining is described in more detail in Chapter 16. 

Subcellular fractionation and immunofluorescence of normal cellular and viral ras proteins have indicated that p21 is localized on the inner face of the plasma membrane. Both the normal and the mutant ras proteins bind GTP specifically and strongly, but only the normal protein has GTPase activity. 

De Rooij KE, Dorsman JC, Smoor MA, Den Dunnen JT, van Ommen GJ. SubceUular localization of the Huntington s disease gene product in cell lines by immunofluorescence and biochemical subcellular fractionation. Hum Mol Genet 1996 5 1093-99. 

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