Abstract
Immunofluorescence microscopy of cultured cells often gives poor preservation of delicate structures. We have obtained dramatically improved results with a simple modification of a standard protocol. Cells growing on a coverslip are rapidly dehydrated in a cold organic solvent and then are rehydrated in a solution containing a homobifunctional crosslinker. The crosslinking reaction stabilizes cellular structures during subsequent incubation and wash steps, usually without compromising antigenicity. This method reproducibly yields high-quality images of endomembrane compartments and cytoskeletal elements.
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Acknowledgments
This work was supported by NIH grant GM-61156. The anti-Sec13 antibody was a kind gift of Bor Luen Tang and Wanjin Hong (National University of Singapore).
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Bhattacharyya, D., Hammond, A.T., Glick, B.S. (2010). High-Quality Immunofluorescence of Cultured Cells. In: Economou, A. (eds) Protein Secretion. Methods in Molecular Biology, vol 619. Humana Press. https://doi.org/10.1007/978-1-60327-412-8_24
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DOI: https://doi.org/10.1007/978-1-60327-412-8_24
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Online ISBN: 978-1-60327-412-8
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