Reporter gene assay β-galactosidase substrates - H9C2

Reporter gene assays enable high sensitivity measurement of gene expression and cell signaling through the addition of bioluminescent genes into target cells. One of the major challenges is to make a specific construct that has no responses other than those related to the signaling pathway of interest. This can be achieved by selecting highly specific reporter constructs containing only defined responsive elements and a minimal promoter linked to reporter enzymes such as luciferase

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Found 3 matching solutions for this experiment

Upstream tips
Pretreat cells with TM5441 for 24 h followed by Doxorubicin (50 nM) treatment in triplicate for 4 days.
Protocol tips
Fix cells with fixative solution for 15 min at room temperature.

Cover the plate with foil and i incubate overnight at 37°C without Co2.
Protocol tips
Fix cells for 15 min at room temperature with fixative solution.

Add 1 ml of the β-Galactosidase Staining Solution and incubate the plate at 37°C at least overnight in a dry incubator (no CO2)
Protocol tips
Add 100µL of β-Galactosidase Assay Reagent.

Cover plate and incubate for 30 minutes at 37°C.
Downstream tips
Measure absorbance at 405nm.
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