Luminescent β-galactosidase Detection Kit II

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Warm enough Reaction Buffer and Reaction Substrate for the entire experiment to room temperature	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

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.

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 - MDA-MB-231 using Luminescent β-galactosidase Detection Kit II from Takara Bio Inc.

Manufacturer protocol

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

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Reporter gene assay β-galactosidase substrates - MDA-MB-231 using Luminescent β-galactosidase Detection Kit II from Takara Bio Inc. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.