Galacto-Light Plus™ β-Galactosidase Reporter Gene Assay System

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed ~ 2 × 105 cells/well	Add diluted Galacton-Plus substrate 1:100 with Reaction Buffer Diluent. Add 70µL of Reaction Buffer, mix, and incubate for 30-60 min

Upstream tips
Seed ~ 2 × 105 cells/well
Protocol tips
Add diluted Galacton-Plus substrate 1:100 with Reaction Buffer Diluent. Add 70µL of Reaction Buffer, mix, and incubate for 30-60 min

Publication protocol

C2C12 cells were grown in a 24-well plate (~ 2 × 105 cells/well) in DMEM supplemented with 10% FBS. After reaching approximately 70% confluence, cells were transfected using Superfect transfection reagent (Qiagen, Hilden, Germany) with 500 ng/well of each specific luciferase construction and 25 ng/well of SV40-β-Gal as an internal control for transfection efficiency. Six hours after transfection, the cells were fed with fresh medium (DMEM with 10% FBS) and incubated overnight. Transfected cells were then treated for 48 h, as indicated in specific experiments. Cell extracts were then prepared and luciferase assays were done using the Luciferase Assay System (Promega, Madison, WI, USA). Luciferase activities were normalized with respect to parallel beta-galactosidase activities, to correct for differences in transfection efficiency. Beta-galactosidase assays were performed using the Galacto-Light Plus System from Tropix (Bedford, MA, USA).

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 - C2C12 using Galacto-Light Plus™ β-Galactosidase Reporter Gene Assay System from Thermo Fisher Scientific.

Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for Galacto-Light Plus™ β-Galactosidase Reporter Gene Assay System below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Reporter gene assay β-galactosidase substrates - C2C12 using Galacto-Light Plus™ β-Galactosidase Reporter Gene Assay System from Thermo Fisher Scientific. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.