Browsing by Author "Singleton, Eileen Willey"

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    A protocol for whole-mount immuno-coupled hybridization chain reaction (WICHCR) in zebrafish embryos and larvae
    (Cell Press, 2021) Ibarra-García-Padilla, Rodrigo; Howard, Aubrey Gaylon Adam; Singleton, Eileen Willey; Uribe, Rosa Anna
    Characterizing mRNA and protein expression with temporal and spatial resolution is a valuable component of nearly every developmental study. Here, we describe a protocol that combines in situ hybridization chain reaction (HCR) and immunofluorescence, allowing for the detection of mRNAs and proteins simultaneously, in zebrafish embryos and larvae. This protocol expands the flexibility of multiplexed HCR by coupling it with traditional immunofluorescence detection. For complete details on the use and execution of this protocol, please refer to Choi et al. (2010, 2016, 2018) and Howard et al. (2021).
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    Determination of Ligand Pathways in Globins: Apolar Tunnels Versus Polar Gates
    (The American Society for Biochemistry and Molecular Biology, Inc., 2012) Salter, Mallory D.; Blouin, George C.; Soman, Jayashree; Singleton, Eileen Willey; Dewilde, Sylvia; Moens, Luc; Pesce, Alessandra; Nardini, Marco; Bolognesi, Martino; Olson, John S.
    Background: O2 pathways in animal hemoglobins and myoglobins are controversial. Results: Ligands enter and exit sperm whale Mb and Cerebratulus lacteus Hb by completely different pathways. Conclusion: Rational mutagenesis mapping can identify ligand migration pathways and provides experimental benchmarks for testing molecular dynamics simulations. Significance: Globins can use either a polar gate or an apolar tunnel for ligand entry.