ONE-Glo™ Luciferase Assay System

Protocol tips

Upstream tips	Protocol tips	Downstream tips
10^4 cells into 96-well plates for 24 hr	Transfecte with 0.2 μg Tax expression plasmid. Performed luciferase by using One Glo system assay after 48 h of transfection. Add 100µl of reagent to the cells grown in 100µl of medium.	Wait at least 3 minutes to allow complete cell lysis and measure in a luminometer

Upstream tips
10^4 cells into 96-well plates for 24 hr
Protocol tips
Transfecte with 0.2 μg Tax expression plasmid. Performed luciferase by using One Glo system assay after 48 h of transfection. Add 100µl of reagent to the cells grown in 100µl of medium.
Downstream tips
Wait at least 3 minutes to allow complete cell lysis and measure in a luminometer

Publication protocol

BHK-EGFP cells were plated and stably transfected with 5 µg linearized pGL4LTRLuc plasmid using the same procedure for the first transfection. Two days after transfection, the cells were cultured in complete medium containing 0.2 mg/ml hygromycin and 1 mg/ml geniticin (G418) (Invivogen, USA) for 3-4 weeks. Stable colonies were isolated by culturing them at limited dilution in wells of a 96-well plate for three cycles. Subsequently several single clones of the G418-resistant colonies were taken under luciferase assay with following procedure. After seeding 104 cells into 96-well plates for 24 hr, BHK-EGFP-HTLV-1Luc cells were transfected with 0.2 μg Tax expression plasmid using Polyfect reagent (Qiagen, 493277090A012 Germany). Transfection was normalaized as previously described. After additional 48 hr incubation, we per-formed luciferase by using One Glo system assay (Promega-Inc.) according to the manufacturer’s instructions onto a Synergy4 luminometer (Biotek, USA). The cells with high constitutive luciferase expression were removed. Furthermore 27 colonies were selected for the lowest background and high Luc expression upon co-culture with 104 Hut-102 cells for 48 hr.

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 luciferase - BHK-21 baby hamster kidney cells using ONE-Glo™ Luciferase Assay System from Promega.

Manufacturer protocol

Download the product protocol from Promega for ONE-Glo™ Luciferase Assay System below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Reporter gene assay luciferase - BHK-21 baby hamster kidney cells using ONE-Glo™ Luciferase Assay System from Promega. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.