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Antibody Staining for Nematodes with Heat-induced Antigen Retrieval (HIAR)

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C. elegans

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2468))

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Immunocytochemistry remains a valuable and necessary tool for biologists working with nematodes, even those nematode model organisms with advanced molecular genetic tools and transgenics. Because of the highly idiosyncratic nature of successful immunostaining procedures, innovations can still be found for this long-established technique. Heat-induced antigen retrieval (HIAR) is well known from other systems, but seems not to have been applied to antibody staining in nematodes. For some antigens, adding HIAR to an established antibody staining protocol for nematodes can reveal strong and reliable staining that without HIAR is poor or completely absent.

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Pristionchus pacificus strains with epitope-tag (FLAG) knock-ins discussed here were made by Hanh Witte & Ralf Sommer (Max Planck Institute for Developmental Biology, Tübingen, Germany), and initially analyzed while CL was a visiting scholar (on sabbatical) in the laboratory of Oliver Hobert (Department of Biological Sciences, HHMI, Columbia University, NY, USA). This technique was also previously described in [8].

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Correspondence to Curtis M. Loer .

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Loer, C.M. (2022). Antibody Staining for Nematodes with Heat-induced Antigen Retrieval (HIAR). In: Haspel, G., Hart, A.C. (eds) C. elegans. Methods in Molecular Biology, vol 2468. Humana, New York, NY.

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  • Print ISBN: 978-1-0716-2180-6

  • Online ISBN: 978-1-0716-2181-3

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