Reporter gene assay β-galactosidase substrates - A549

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
Plate 3×104/well.
Protocol tips
After 7 days of 100 nM doxorubicin treatement, add 1 ml of the Staining Mixture per well and incubate at 37 °C without CO2 until the cells are stained blue.
Protocol tips
Add 150µl of Assay 2X Buffer to sample.

Incubate the reactions at 37°C for 30 minutes or until a faint yellow color has developed.
Downstream tips
Read the absorbance at 420nm.
Protocol tips
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)
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