β-Gal Reporter Gene Assay, chemiluminescent

Reporter gene assay β-galactosidase substrates - SH-SY5Y

Experiment
Reporter gene assay β-galactosidase substrates - SH-SY5Y
Product
β-Gal Reporter Gene Assay, chemiluminescent from Sigma-Aldrich
Manufacturer
Sigma-Aldrich

Protocol tips

Upstream tips
Culture cells for 48 h
Protocol tips
Wash cells and add β-Gal Reporter Gene Assay, incubate for 30 min at RT.

Add substrate reagent and incubate for 15-60 min at RT

Publication protocol

The basal promoter activity of the different reporter plasmids was assayed by measuring the luciferase activity after transient transfection in HeLa, SH-SY5Y and Ntera2/clone D1 (NT2) cell lines.

To minimize variations in transfection efficiency, replicates were transfected in single batch suspension with FuGENE® HD (Roche Diagnostics GmbH, Penzberg, Germany) according to the manufacturer's instructions. Plates containing 200,000 cells were co-transfected with 0.5 µg of the reporter plasmid together with an expression plasmid containing the β-galactosidase gene-coding region (pSV40-βGAL). Cells were inoculated in 24-well plates and maintained for 48 h. These cells were harvested and lysed in reporter lysis buffer (Promega). Cell extracts were assayed for luciferase and β-galactosidase activity (β-Gal Reporter Gene Assay, Roche), which was used to normalize the results. All experiments were performed at least three times in duplicate well.

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Papers

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Paper title
Demethylation of the Coding Region Triggers the Activation of the Human Testis-Specific Gene in Somatic Tissues
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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for β-Gal Reporter Gene Assay, chemiluminescent below.

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