Reporter gene assay β-galactosidase substrates - HUVEC

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 2×10^5 cells
Protocol tips
co-incubate with dexmedetomidine and LPS for 48 h and then assay cells for Luminescense.

Add GLuc Assay Working Solution and incubate at room temperature for 1 minute
Downstream tips
Read the plate within 5 min after the Incubation
Protocol tips
Fix the cells with 0.5 ml of Fixative Solution for 10-15 minutes at room temperature.

Add 0.5 ml of the Staining Solution Mix.

Cover the plate. Incubate overnight at 37°C
Protocol tips
Add 150µl of Assay 2X Buffer to sample.

Incubate the reactions at 37°C for 30 minutes or until a faint yellow color has developed.
Downstream tips
Read the absorbance at 420nm.
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