β-Gal Reporter Gene Assay, chemiluminescent

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Culture cells for 48 h	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

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.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Reporter gene assay β-galactosidase substrates - SH-SY5Y using β-Gal Reporter Gene Assay, chemiluminescent from Sigma-Aldrich.

Manufacturer protocol

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

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Reporter gene assay β-galactosidase substrates - SH-SY5Y using β-Gal Reporter Gene Assay, chemiluminescent from Sigma-Aldrich. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.