Luminescent β-galactosidase Detection Kit II

Reporter gene assay β-galactosidase substrates - MDA-MB-231

Experiment
Reporter gene assay β-galactosidase substrates - MDA-MB-231
Product
Luminescent β-galactosidase Detection Kit II from Takara Bio Inc
Manufacturer
Takara Bio Inc

Protocol tips

Upstream tips
Warm enough Reaction Buffer and Reaction Substrate for the entire experiment to room temperature
Protocol tips
Add 200 µl of the Reaction Buffer Mixture to each cell lysate and mix gently.

Incubate at room temperature (20–25°C) for 60 min

Publication protocol

Vectors encoding human C/EBPα or β (SC303472 and SC319561; Origene, Burlington, ON) and vectors encoding C/EBPβ-2 and C/EBPβ-3 (No. 15738 and No. 15737; Addgene, Cambridge, MA) were obtained commercially. The cDNA encoding the human galectin-7 (provided by Dr. T. Magnaldo) was cloned in the srα eukaryotic expression vector (kind gift of Dr. François Denis) using SpeI and BamHI restriction sites. Cells were transfected using the Lipofectamine 2000 reagent (Life Technologies) according to the manufacturer's protocol. For reporter assays, NF-κB or a C/EBP luciferase reporter vectors (Cat. No. 219078 and 240112 respectively; Stratagene, Santa Clara, CA) were used. The (empty) pCMV5 vector was used as a control. Transfection efficiency was measured using the pCMV/β-gal reporter vector (Promega, Madison, WI). Luciferase activity was measured using the Luciferase Assay System protocol (Promega) and a luminometer (Lumat LB 9507, Berthold). β-galactosidase activity was detected using a colorimetric enzyme assay using the Luminescent β-Galactosidase Detection Kit II according to the manufacturer's instructions (Clontech Laboratories, Mountain View, CA). Luciferase expression levels were normalized to the levels of β-galactosidase expression.

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Reporter gene assay β-galactosidase substrates - MDA-MB-231 using Luminescent β-galactosidase Detection Kit II from Takara Bio Inc.

Paper title
The CCAAT/Enhancer-Binding Protein Beta-2 Isoform (CEBPβ-2) Upregulates Galectin-7 Expression in Human Breast Cancer Cells
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Manufacturer protocol

Download the product protocol from Takara Bio Inc for Luminescent β-galactosidase Detection Kit II below.

Download PDF Download manufacturer protocol

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