Pcmv renilla luciferase

Manufactured by Promega

Sourced in France

The PCMV-Renilla luciferase is a lab equipment product designed to measure the expression of the Renilla luciferase reporter gene. It consists of a plasmid vector that contains the Renilla luciferase gene under the control of the cytomegalovirus (CMV) promoter.

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Lab products found in correlation

Dual glo assay system, by Promega (2 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) 8xgtiic luciferase, by Addgene (1 mentions) Pcdna3 vector, by Thermo Fisher Scientific (1 mentions) Quikchange lightning multi site directed mutagenesis kit, by Agilent Technologies (1 mentions) Dual luciferase assay, by Promega (1 mentions) Passive lysis buffer (plb), by Promega (1 mentions)

4 protocols using pcmv renilla luciferase

Exploring autophagy regulation by α-catenin

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The following constructs were used in this study: 8XGTIIC-luciferase (no. 34615, Addgene)^{32 (link)}, pCMV-Renilla Luciferase (E2261, Promega), empty mEmerald-C1 (no. 54734, Addgene), mEmerald-Alpha1-Catenin-C-18 (no. 53982, Addgene), GFP-YAP (no. 17843, Addgene). The mRFP-LC3 construct was kindly provided by Dr. Tamotsu Yoshimori (Osaka University, Japan). Alpha1-Catenin which was amplified from mEmerald-Alpha1-Catenin-C-18 was cloned into pFlag-CMV-5a expression vector (E7523, Sigma-Aldrich) using EcoRI and SalI restriction enzyme sites (pFlag-CTNNA1).
Pre-designed siRNAs (On-Target plus SMART pool and/or set of deconvoluted oligos) targeting Control (D-001810-10), ATG7 (L-020112-00), ATG10 (L-019426-01), ATG16L1 (L-021033-01), CTNNA1 (L-010505-00) and CTNNA3 (L-020300-00) were purchased from Dharmacon Thermo Scientific.
(1) set of deconvoluted oligos for ATG7/10 #1 (J-020112-05, J-019426-09):
ATG7 siRNA_05: CCAACACACUCGAGUCUUU
ATG10 siRNA_09: CGUCUCAGGAUGAACGAAA
(2) set of deconvoluted oligos for ATG7/10 #2 (J-020112-06, J-019426-10):
ATG7 siRNA_06: GAUCUAAAUCUCAAACUGA
ATG10 siRNA_10: AGGAAUUGCGGCACGAAGA
(3) set of deconvoluted oligos for ATG7/10 #3 (J-020112-06, J-019426-11):
ATG7 siRNA_06: GAUCUAAAUCUCAAACUGA
ATG10 siRNA_11: GGAGGAGGCUUUCGAGCUA
(4) set of deconvoluted oligos for ATG7/10 #4 (J-020112-06, J-019426-12):
ATG7 siRNA_06: GAUCUAAAUCUCAAACUGA
ATG10 siRNA_12: CCAACGUUAUUGUGCAGAA

Pavel M., Park S.J., Frake R.A., Son S.M., Manni M.M., Bento C.F., Renna M., Ricketts T., Menzies F.M., Tanasa R, & Rubinsztein D.C. (2021). α-Catenin levels determine direction of YAP/TAZ response to autophagy perturbation. Nature Communications, 12, 1703.

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Wnt Signaling Activation Assay

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MC3T3-E1 cells were co-transfected with 400 ng TOPflash-luc reporter plasmid and 10 ng control pCMV-Renilla luciferase (Promega) using Lipofectamine 2000 (Invitrogen) according to the manufacturer’s protocol. Six hours after transfection, the medium was changed, and cells were cultured in α-MEM containing 1% FBS overnight. Cells were then treated with WNT16b (0.2 μg ml⁻¹) or WNT3a (0.2 μg ml⁻¹) for 24 h, followed by luciferase assay using the Dual-Glo Assay system (Promega) according to the manufacturer’s instructions. Data were normalized by Renilla firefly activity and are presented as a fold change compare to vehicle-treated cells. HEK293 cells stably expressing ROR2-hTRK-luc were incubated with WNT16 (0.2 μg ml⁻¹) or WNT5a (0.2 μg ml⁻¹) for 24 h. Luciferase activities were determined as described above.

Movérare-Skrtic S., Henning P., Liu X., Nagano K., Saito H., Börjesson A.E., Sjögren K., Windahl S.H., Farman H., Kindlund B., Engdahl C., Koskela A., Zhang F.P., Eriksson E.E., Zaman F., Hammarstedt A., Isaksson H., Bally M., Kassem A., Lindholm C., Sandberg O., Aspenberg P., Sävendahl L., Feng J.Q., Tuckermann J., Tuukkanen J., Poutanen M., Baron R., Lerner U.H., Gori F, & Ohlsson C. (2014). Osteoblast-derived WNT16 represses osteoclastogenesis and prevents cortical bone fragility fractures. Nature medicine, 20(11), 1279-1288.

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LRP5 Signaling Pathway Activation

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The wild‐type construct of the full‐length LRP5 expression plasmid, cloned into the pcDNA3 vector (Invitrogen), was kindly provided by Dr. M. Warman (Harvard Medical School, Boston, MA, USA). The site‐directed mutagenesis involved use of the QuikChange Lightning Multi Site‐Directed Mutagenesis Kit (Agilent). Saos‐2 cells were co‐transfected by using Lipofectamine 2000 (Invitrogen, Paris, France) with the TOPflash‐luc reporter plasmid, pCMV‐Renilla luciferase (Promega, Charbonniere‐les‐bains, France) and an expression‐mutated or control plasmid (expression plasmid without cDNA of LRP5 and expression plasmid with wild‐type cDNA of LRP5). Saos‐2 cells were transfected only with TOPflash‐luc reporter plasmid, pCMV‐Renilla luciferase (a transcription factor activated by Wnt signaling). All experiments were realized in triplicate. Cells were plated at 1.25 × 105 cells/well in 24‐well plates for transfection. Cells were cultured in Wnt3a‐conditioned medium or not for 48 hours before transfection.18 The luciferase assay involved the Dual‐Glo Assay system (Promega) according to the manufacturer's instructions. Data were normalized by Renilla firefly activity and are presented as a fold change compared to vehicle‐treated cells.

Collet C., Ostertag A., Ricquebourg M., Delecourt M., Tueur G., Isidor B., Guillot P., Schaefer E., Javier R., Funck‐Brentano T., Orcel P., Laplanche J, & Cohen‐Solal M. (2017). Primary Osteoporosis in Young Adults: Genetic Basis and Identification of Novel Variants in Causal Genes. JBMR Plus, 2(1), 12-21.

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BACE1 Promoter Regulation Assay

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The human BACE1 promoter region from −2000 to −1 was amplified by PCR using the primers XhoI site (5′-CCGCTCGAGATTCTATTTTTCCTGTAGTTTTATT-3ʹ) and −1 Bg1III site (5′-GAAGATCTGGCGGCGGCTGTCAAAGCCAAAAGG-3ʹ) from the human SH-SY5Y cell genome. Nucleotides in bold represent restriction enzyme cutting sites. The PCR product was cloned in front of the firefly luciferase gene of pGL3-basic vector at XhoI and Bg1III site to generate pBACE1-Luci reporter plasmid. The plasmid was further confirmed by DNA sequencing.
293APPswe cells were cultured in black 96-well plates and grown to approximately 70% confluence, then transfected with 0.2 μg plasmid/100 μl per well using lipo-LTX reagent (Invitrogen) according to the manufacturer’s protocol. pCMV-Renilla luciferase (Promega) was co-transfected in order to normalize transfection efficiency. At 24 hours after transfection, 25 μl of cell medium per well was replaced by 75 μl of passive lysis buffer (Promega), and cells were incubated for 10 minutes at room temperature to allow cells to lyse. The firefly luciferase activity and Renilla luciferase activity assays were performed sequentially in one well with a luminometer (PerkinElmer), according to the instructions for the Dual-luciferase assay (Promega). The firefly luciferase activity was normalized to the Renilla luciferase activity and expressed as relative luciferase units (RLU).

Wang Q., Xiao B., Cui S., Song H., Qian Y., Dong L., An H., Cui Y., Zhang W., He Y., Zhang J., Yang J., Zhang F., Hu G., Gong X., Yan Z., Zheng Y, & Wang X. (2014). Triptolide treatment reduces Alzheimer’s disease (AD)-like pathology through inhibition of BACE1 in a transgenic mouse model of AD. Disease Models & Mechanisms, 7(12), 1385-1395.

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