Luciferase Assay for Zebrafish Larvae

Embryos were microinjected as described above with 200 pg of in vitro-transcribed zfiGLuc or GLuc RNA. After 48 h, larvae were infected with WT ST (MOI of 10) and whole-larval extracts were obtained at 24 hpi as described previously65 (link). Larval extracts were then combined 1:1 with distilled water containing 4.4 μM coelenterazine (Sigma-Aldrich) to achieve a final concentration of 2.2 μM. The luciferase signal was then measured on a Luminometer Optocomp II (MGM Instruments).

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Tyrkalska S.D., Candel S., Angosto D., Gómez-Abellán V., Martín-Sánchez F., García-Moreno D., Zapata-Pérez R., Sánchez-Ferrer Á., Sepulcre M.P., Pelegrín P, & Mulero V. (2016). Neutrophils mediate Salmonella Typhimurium clearance through the GBP4 inflammasome-dependent production of prostaglandins. Nature Communications, 7, 12077.

Publication 2016

coelenterazine

Coelenterazine Embryos Larval Luciferase

Corresponding Organization :

Other organizations : Universidad de Murcia, Hospital Universitario Virgen de la Arrixaca, Centro de Investigación Biomédica en Red

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Variable analysis

independent variables

Microinjection of 200 pg of in vitro-transcribed zfiGLuc or GLuc RNA into embryos

dependent variables

Luciferase signal measured on a Luminometer Optocomp II

control variables

Embryos were microinjected
Larvae were infected with WT ST (MOI of 10)
Larval extracts were obtained at 24 hpi
Larval extracts were combined 1:1 with distilled water containing 4.4 μM coelenterazine

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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