[14]Xenopus oocyte microinjection: From gene to protein
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2023, Pesticide Biochemistry and PhysiologyCitation Excerpt :Following anesthesia in 1.0 L of MS-222 mixture (2 g of ethyl 3-aminobenzoate methansulfonate salt and 0.5 g of sodium bicarbonate (NaHCO3)), oocytes were surgically removed from female frogs and defollicated by incubation in OR2 buffer (82.5 mM NaCl, 2 mM KCl, 1 mM MgCl2, 5 mM HEPES, pH = 7.4) supplemented with Type 1A collagenase (2 mg/ml) purchased from Sigma-Aldrich, Corp. (St. Louis, MO) and the remaining follicle membrane removed manually following treatment (Marsal et al., 1995; Miledi et al., 2002). Isolated stage V–VI oocytes were incubated in ND96 medium (96 mM NaCl, 2 mM KCl, 1.8 mM CaCl2, 1 mM MgCl2, and 5 mM HEPES, pH 7.4) supplemented with 1% sodium pyruvate, 1% penicillin/streptomycin, and 5% horse serum overnight at 19 °C prior to injection (Soreq, 1992). Surgery was performed in accordance to IACUC protocol ID # 2013–0026.
Evaluation of microtransplantation of rat brain neurolemma into Xenopus laevis oocytes as a technique to study the effect of neurotoxicants on endogenous voltage-sensitive ion channels
2017, NeuroToxicologyCitation Excerpt :Following anesthesia in 1.0 l of MS-222 mixture (2 g of ethyl 3-aminobenzoate methansulfonate salt and 0.5 g of sodium bicarbonate (NaHCO3)), oocytes were surgically removed from female frogs and defollicated by incubation in OR2 buffer (82.5 mM NaCl, 2 mM KCl, 1 mM MgCl2, 5 mM HEPES, pH = 7.4) supplemented with Type 1A collagenase (2 mg/ml) purchased from Sigma-Aldrich, Corp. (St. Louis, MO) and the remaining follicle membrane removed manually following treatment (Marsal et al., 1995; Miledi et al., 2002). Isolated stage V–VI oocytes were incubated in ND96 medium (96 mM NaCl, 2 mM KCl, 1.8 mM CaCl2, 1 mM MgCl2, and 5 mM HEPES, pH 7.4) supplemented with 1% sodium pyruvate, 1% penicillin/streptomycin, and 5% horse serum overnight at 19 °C prior to injection (Soreq, 1992). Surgery was performed in accordance to IAUCUC protocol ID # 2013-0026.
Intestinal dehydroascorbic acid (DHA) transport mediated by the facilitative sugar transporters, GLUT2 and GLUT8
2013, Journal of Biological ChemistryCitation Excerpt :All cDNA sequences were verified by automated DNA sequencing. Oocytes were isolated from Xenopus laevis and injected with mRNA using established methods (18). Briefly, mature adult female frogs were anesthetized with 3-aminobenzoic acid ethyl ester (2 g/750 ml) in ice water.
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