out brain areas.
small plunger to homogenize tissue samples directly in a polyethylene tube.
Eppendorf tubes to pellet cell debris after homogenization so as to recuperate supernatant for luciferase assay.
This system is based on the oxidation of luciferin by luciferase, in the presence of ATP and O2, with photon production.
Tropix, Bedford, MA, USA) to quantify light emitted.
To quantify luciferase expression, animals are sacrificed by decapitation after anesthesia. The dissected brains are separated in hemispheres and homogenized using an Ultra-Turrax, in 2-mL Eppendorf tubes containing 200 μL (for the newborn) or 500 μL (for the adults) of luciferase lysis buffer. Homogenates are centrifuged, and 20 μL of each supernatant with 100 μL of luciferase assay substrate are vortex mixed. The light emitted is quantified in relative light units (RLUs) using a luminometer.
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