Abstract
Antibody-based detection of protein distribution patterns both in wholemount and on sections revolutionized Xenopus research and ushered in the visual-based era of Xenopus data presentation. The ability to view the distribution of a gene product throughout an embryo makes it possible to rapidly map normal expression profiles and profiles that have been altered by an experimental intervention. The main limiting element in Xenopus immunostaining techniques has always been the availability of antibodies that work well on fixed whole embryos, a problem that persists. However, new antibodies are constantly being generated and improvements in detection systems allow antibodies that were once below the limits of detection to be utilized in multichannel immunofluorescence using tyramide amplification.
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References
Dent JA, Polson AG, Klymkowsky MW (1989) A whole-mount immunocytochemical analysis of the expression of the intermediate filament protein vimentin in Xenopus. Development 105:61–74
Klymkowsky MW, Hanken J (1991) Whole-mount staining of Xenopus and other vertebrates. Methods Cell Biol 36:419–441
Hemmati-Brivanlou A, Melton DA (1992) A truncated activin receptor inhibits mesoderm induction and formation of axial structures in Xenopus embryos. Nature 359:609–614
Saka Y, Smith JC (2001) Spatial and temporal patterns of cell division during early Xenopus embryogenesis. Dev Biol 229:307–318
Faure S, Lee MA, Keller T, ten Dijke P, Whitman M (2000) Endogenous patterns of TGFbeta superfamily signaling during early Xenopus development. Development 127:2917–2931
Dubaissi E, Papalopulu N (2011) Embryonic frog epidermis: a model for the study of cell-cell interactions in the development of mucociliary disease. Dis Model Mech 4:179–192
Vize PD, Melton DA, Hemmati-Brivanlou A, Harland RM (1991) Assays for gene function in developing Xenopus embryos. Methods Cell Biol 36:367–387
Kroll KL, Amaya E (1996) Transgenic Xenopus embryos from sperm nuclear transplantations reveal FGF signaling requirements during gastrulation. Development 122:3173–3183
Drummond IA, Majumdar A, Hentschel H, Elger M, Solnica-Krezel L, Schier AF et al (1998) Early development of the zebrafish pronephros and analysis of mutations affecting pronephric function. Development 125:4655–4667
Kapust RB, Waugh DS (1999) Escherichia coli maltose-binding protein is uncommonly effective at promoting the solubility of polypeptides to which it is fused. Protein Sci 8:1668–1674
Fox JD, Waugh DS (2003) Maltose-binding protein as a solubility enhancer. Methods Mol Biol 205:99–117
Acknowledgements
This work was supported by the Wellcome Trust to NP and NIH R01 HD45776 and P41 HD064556 to PDV. The authors would like to thank Xiaolan Zhou for his role in developing tyramide protocols.
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Dubaissi, E., Panagiotaki, N., Papalopulu, N., Vize, P.D. (2012). Antibody Development and Use in Chromogenic and Fluorescent Immunostaining. In: HOPPLER, S., Vize, P. (eds) Xenopus Protocols. Methods in Molecular Biology, vol 917. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-992-1_23
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DOI: https://doi.org/10.1007/978-1-61779-992-1_23
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