Reporter gene assay β-galactosidase substrates - mouse mesenchymal stem 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 no responses other than those related to the signaling pathway of interest. This can be achieved by selecting a highly specific reporter constructs containing only defined responsive elements and a minimal promoter linked to reporter enzyme such as luciferase.

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Found 3 matching solutions for this experiment

Upstream tips
Seed cells for 24 hours and treat with TGF-β1 for another 24 hours.
Protocol tips
Fix cells with 0.5 mL 1Х fixative solution for 15 min at room temperature.

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)
Upstream tips
Pretreat cells with LPS.

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 for 15 minutes in a fixative solution

Add 1 ml of the Staining Mixture per well and incubate overnight at 37 °C without CO2 until the cells are stained blue
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