Reporter gene assay β-galactosidase substrates - U2OS

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

Protocol tips
Dilute Galacton-Plus substrate 1:100 in Buffer B.

Add 25 μL of Buffer A to each well.

Within 10 min, inject 100 μL of Buffer B (containing Galacton-Plus substrate). After a 1-2 sec delay, read the luciferase signal for 0.1-1 sec/well

Incubate for 30-60 min at room temperature
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
Protocol tips
Fix cells with with 2% formaldehyde/0.2% glutaraldehyde.

Incubate for 12 hours at 37 ° C without Co2
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