Reporter gene assay β-galactosidase substrates - Aspc-1

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
Seed cells fro 72 hours and washe twice with PBS,

Fix for 6 min with fixation buffer containing 2% formaldehyde, 0.2% glutaraldehyde, 7.0 mM Na2HPO4, 1.5 mM KH2PO4, 140 mM NaCl, and 2.68 mM KCl
Protocol tips
Add 1 ml of the Staining Mixture per well and incubate overnight at 37 °C without CO2 until the cells are stained blue.
Upstream tips
Fix cells in 1x fixation solution containing 2% formaldehyde and 0.2% glutaraldehyde in PBS
Protocol tips
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
Seed 5 × 10^4 cells
Protocol tips
Add 100µL of β-Galactosidase Assay Reagent.

Cover plate and incubate for 30 minutes at 37°C.
Downstream tips
Measure absorbance at 405nm.
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