Reporter gene assay β-galactosidase substrates - C2C12

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
Add Fixing Buffer to the dish and incubate for 10-15 minutes at room temperature.

Add 1X X-Gal staining solution and incubate the cells between 1-18 hours at 37°C in a humidified incubator.
Protocol tips
Remove medium and rinse cells in 1X PBS.

Add 1X lysis reagent.

Mix 20µl of cell lysate with 100µl of Luciferase Assay Reagent and measure the light produced.
Upstream tips
Seed ~ 2 × 105 cells/well
Protocol tips
Add diluted Galacton-Plus substrate 1:100 with Reaction Buffer Diluent.

Add 70µL of Reaction Buffer, mix, and incubate for 30-60 min
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