Reporter gene assay β-galactosidase substrates - H1299

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
Warm enough Reaction Buffer and Reaction Substrate for the entire experiment to room temperature
Protocol tips
Add 200 µl of the Reaction Buffer Mixture to each cell lysate and mix gently.

Incubate at room temperature (20–25°C) for 60 min
Upstream tips
Seed 3 × 10^5 cells
Protocol tips
After 24 hours of transfection, perform assay and incubate for 30 min at 37 °C or until a faint yellow colour has developed.
Downstream tips
Read the absorbance at 420nm
Protocol tips
Treat cells with kit after 96 hrs of incubation.

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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