Reporter gene assay β-galactosidase substrates - HeLa cervical cancer cells

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
Analyze the β-galactosidase expression 40-72 hr

Add lysis buffer and incubate at room temperature for 15 minutes.

Add 2x assay buffer and incubate at 37 °C for 30 minutes or until a yellow color develops (t min).
Downstream tips
Read OD at 420 nm
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 cells for 48hours
Protocol tips
Fix cells with 4% formaldehyde.

Incubate overnight at 37 ° C without Co2.
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