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Prl sv40 renilla luciferase reporter

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The PRL-SV40 Renilla luciferase reporter is a lab equipment product used to measure gene expression. It contains the Renilla luciferase reporter gene under the control of the SV40 promoter. Renilla luciferase is a naturally occurring enzyme that produces light when exposed to its substrate, coelenterazine. The PRL-SV40 reporter can be used to quantify gene expression levels in various cell lines and experimental systems.

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Lab products found in correlation

Lipofectamine 2000, by Thermo Fisher Scientific (10 mentions) Dual luciferase reporter assay system, by Promega (5 mentions) Dual luciferase reporter assay, by Promega (4 mentions) P3 primary cell nucleofector kit, by Lonza (2 mentions) Soluble anti cd28 cd28.2, by BD (2 mentions) Human il 2, by Thermo Fisher Scientific (2 mentions) Anti cd3 cd3 2, by Mabtech (2 mentions) Ap 1 luciferase reporter plasmid, by Addgene (2 mentions) Amaxa nucleofector system, by Lonza (2 mentions) Glomax system, by Promega (2 mentions) Dual glo luciferase reporter kit, by Promega (2 mentions) Pgl3 basic luciferase reporter vector, by Promega (2 mentions) Lipofectamine ltx, by Thermo Fisher Scientific (2 mentions) Snail 3 utr firefly luciferase reporter vector, by Promega (2 mentions) Pgl3 control vector, by Promega (1 mentions)

Spelling variants of this product

PRL-SV40 (378 citations) Renilla luciferase (170 citations) Renilla luciferase reporter (32 citations) PRL-SV40 Renilla luciferase (11 citations) SV40-Renilla (11 citations) PRL-SV40 Renilla luciferase reporter vector (10 citations) PRL-SV40 Renilla luciferase reporter plasmids (8 citations) PRL-SV40 Renilla luciferase control reporter vector (6 citations) Renilla reporter pRL-SV40 (4 citations) Renilla-luciferase (pRL) (2 citations)

11 protocols using prl sv40 renilla luciferase reporter

1

Transient Transfection and UVB Treatment

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HaCaT cells were grown to ~60% confluency in 3.5 cm plates and transfected with the indicated plasmids and concentrations using Lipofectamine 2000 as per the manufacturer’s instructions (ThermoFisher). This includes 1 μg of either the pGL3-control vector (Promega) or the pGL3-FHRE Firefly luciferase reporters co-transfected with 0.1 μg of the pRL-SV40 Renilla luciferase reporter (Promega) for the luciferase assay. UVB treatment was performed 48 h following transfection.
Feehan R.P., Nelson A.M, & Shantz L.M. (2018). Inhibition of mTORC2 enhances UVB-induced apoptosis in keratinocytes through a mechanism dependent on the FOXO3a transcriptional target NOXA but independent of TRAIL. Cellular signalling, 52, 35-47.
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2

T Cell Activation and Transcriptional Assay

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Naive CD4+ T cells were activated with anti-CD3/anti-CD28 beads and transduced with a lentiviral vector as described above. After 36 hours, activated cells were cotransfected with the AP-1 luciferase reporter plasmid (Plasmid #40342; Addgene) along with pRL-SV40 renilla luciferase reporter (Promega) using the Amaxa Nucleofector system and P3 primary cell Nucleofector Kit (Lonza). After 4 hours, activated cells were cultured on plates coated with 1 μg/ml anti-CD3 (CD3–2; Mabtech) plus 2 mg/ml soluble anti-CD28 (CD28.2; BD Biosciences) and 10 U/ml human IL-2 (Peprotech). On day 3 after activation, cells were lysed and luciferase reporter activity was measured with the Dual-Luciferase Reporter Assay System (Promega).
Kim C., Hu B., Jadhav R.R., Jin J., Zhang H., Cavanagh M.M., Akondy R.S., Ahmed R., Weyand C.M, & Goronzy J.J. (2018). Activation of miR-21-Regulated Pathways in Immune Aging Selects against Signatures Characteristic of Memory T Cells. Cell reports, 25(8), 2148-2162.e5.
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3

Transcriptional Regulation of IL10R1

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The 5′ flanking promoter sequence of the IL10R1 gene (from −994 to +23 bp) was amplified from human genomic DNA and subcloned into the pGL3–basic luciferase reporter vector (Promega) to generate pIL10R1-Luc. Caco-2 cells were seeded into 24-well dishes and transfected with the luciferase reporter plasmid pIL10R1-Luc or empty pGL3-basic-Luc (100 ng) using Lipofectamine-LTX (Invitrogen). To assay promoter activity, cells were exposed to 1 μM FICZ, 100 μM Kyn, or 10 ng/mL IFN- γ 24 h posttransfection and harvested at 48 h posttransfection and luciferase activity was measured in extracts using the Dual-Glo luciferase reporter kit and GloMax system (Promega). All firefly luciferase activity was normalized to a cotransfected pRL-SV40 Renilla luciferase reporter (Promega).
Lanis J.M., Alexeev E.E., Curtis V.F., Kitzenberg D.A., Kao D.J., Battista K.D., Gerich M.E., Glover L.E., Kominsky D.J, & Colgan S.P. (2017). Tryptophan Metabolite Activation of the Aryl Hydrocarbon Receptor Regulates IL10 Receptor Expression on Intestinal Epithelia. Mucosal immunology, 10(5), 1133-1144.
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4

Transcriptional Regulation by cMyc-STAT3

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293T cells in 24-well plates (1 × 105 cells per well) were cotransfected with 0.5 μg of firefly luciferase reporter, 0.02 μg of pRL-SV40 renilla luciferase reporter (Promega) and wild-type or mutant cMyc-STAT3 using Lipofectamine 2000 according to the manufacturer's instruction (Life Technologies) for 24 h followed by treatment with IL-6 (20 ng/ml), LPS (10 μg/ml), LIF (20 ng/ml) or no treatment. Twenty-four hours later, whole cell lysates were prepared for the luciferase reporter activity analysis using a Dual-Luciferase® Reporter Assay System (Promega). The pRL-SV40 plasmid was used to normalize transfection efficiency. Data are presented as the mean ± SD of three experiments.
Xu L., Ji J.J., Le W., Xu Y.S., Dou D., Pan J., Jiao Y., Zhong T., Wu D., Wang Y., Wen C., Xie G.Q., Yao F., Zhao H., Fan Y.S, & Chin Y.E. (2015). The STAT3 HIES mutation is a gain-of-function mutation that activates genes via AGG-element carrying promoters. Nucleic Acids Research, 43(18), 8898-8912.
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5

T Cell Activation and Transcriptional Assay

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Naive CD4+ T cells were activated with anti-CD3/anti-CD28 beads and transduced with a lentiviral vector as described above. After 36 hours, activated cells were cotransfected with the AP-1 luciferase reporter plasmid (Plasmid #40342; Addgene) along with pRL-SV40 renilla luciferase reporter (Promega) using the Amaxa Nucleofector system and P3 primary cell Nucleofector Kit (Lonza). After 4 hours, activated cells were cultured on plates coated with 1 μg/ml anti-CD3 (CD3–2; Mabtech) plus 2 mg/ml soluble anti-CD28 (CD28.2; BD Biosciences) and 10 U/ml human IL-2 (Peprotech). On day 3 after activation, cells were lysed and luciferase reporter activity was measured with the Dual-Luciferase Reporter Assay System (Promega).
Kim C., Hu B., Jadhav R.R., Jin J., Zhang H., Cavanagh M.M., Akondy R.S., Ahmed R., Weyand C.M, & Goronzy J.J. (2018). Activation of miR-21-Regulated Pathways in Immune Aging Selects against Signatures Characteristic of Memory T Cells. Cell reports, 25(8), 2148-2162.e5.
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6

Luciferase Assays for Ketamine and HNK Effects

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U251-MG cells were used to perform luciferase reporter gene assays. Cells (1×105) were transfected using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) with 1 μg of the plasmid DNA and 100 ng of pRL-SV40 Renilla luciferase reporter as an internal control (Promega, Madion, WI, USA) in a 6-well plate. After 24 hours, the cells were treated with increasing concentrations of ketamine, (2R,6R)-HNK or (2S,6S)-HNK for 24 hours. Luciferase assays were performed using a dual-luciferase reporter assay system (Promega, Madison, WI, USA). Renilla activity was used to correct for possible variation in transfection efficiency. The pGL3 basic vector without insert was used as a negative control. Expression was calculated as the relative firefly luciferase activity normalized by use of the activity of the transfection control, Renilla luciferase. Results are presented as fold change in relative luciferase units as compared to the pGL3 basic vector. All experiments were repeated three times in triplicate.
Ho M.F., Correia C., Ingle J.N., Kaddurah-Daouk R., Wang L., Kaufmann S.H, & Weinshilboum R.M. (2018). Ketamine and Ketamine Metabolites as Novel Estrogen Receptor Ligands: Induction of Cytochrome P450 and AMPA Glutamate Receptor Gene Expression. Biochemical pharmacology, 152, 279-292.
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7

Transcriptional Regulation of IL10R1

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The 5′ flanking promoter sequence of the IL10R1 gene (from −994 to +23 bp) was amplified from human genomic DNA and subcloned into the pGL3–basic luciferase reporter vector (Promega) to generate pIL10R1-Luc. Caco-2 cells were seeded into 24-well dishes and transfected with the luciferase reporter plasmid pIL10R1-Luc or empty pGL3-basic-Luc (100 ng) using Lipofectamine-LTX (Invitrogen). To assay promoter activity, cells were exposed to 1 μM FICZ, 100 μM Kyn, or 10 ng/mL IFN- γ 24 h posttransfection and harvested at 48 h posttransfection and luciferase activity was measured in extracts using the Dual-Glo luciferase reporter kit and GloMax system (Promega). All firefly luciferase activity was normalized to a cotransfected pRL-SV40 Renilla luciferase reporter (Promega).
Lanis J.M., Alexeev E.E., Curtis V.F., Kitzenberg D.A., Kao D.J., Battista K.D., Gerich M.E., Glover L.E., Kominsky D.J, & Colgan S.P. (2017). Tryptophan Metabolite Activation of the Aryl Hydrocarbon Receptor Regulates IL10 Receptor Expression on Intestinal Epithelia. Mucosal immunology, 10(5), 1133-1144.
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8

Measuring MYC Promoter Activity

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The relative activity of MYC promoter was determined by Dual‐Luciferase reporter assay as previously described.[52] MYC promoter was amplified from genomic DNA of 293T cells by polymerase chain reaction (PCR) and cloned into luciferase reporter construct pGL4.10 (Promega) and confirmed by sequencing. Considering that TAF10 is a cofactor of the transcription factor IID (TFIID) complexes, which recognise the conserved TATA‐box to position the polymerase properly, the native TATA box sequence TATATAAA was mutated into GGTCTGC on the MYC promoter by overlap extension PCR and defined as pGL4.10‐MYC‐MUT. MCF7, A549 and HepG2 cells were seeded in 24‐well plates and transfected with pGL4.10‐MYC or pGL4.10‐MYC‐MUT plasmids by VigoFect (Vigorous Biotech, Beijing). Luciferase activities were measured 24 h after transfection using the Dual‐Luciferase reporter assay system (Promega). The pRL‐SV40 renilla luciferase reporter (Promega) was included in all transfections for normalisation.
Xiong Y., Wang L., Xu S., Fu B., Che Y., Zaky M.Y., Tian R., Yao R., Guo D., Sha Z., Lin F., Lin X, & Wu H. (2023). Small molecule Z363 co‐regulates TAF10 and MYC via the E3 ligase TRIP12 to suppress tumour growth. Clinical and Translational Medicine, 13(1), e1153.
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9

miR-622 Regulation of HULC Expression

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HULC, containing the miR-622 binding site sequence, was cloned into the firefly luciferase reporter vector. The vector and an empty vector were purchased from OriGene Technologies (Rockville, MD, USA). Cells (2 × 105 per well) were plated into 6-well plates and treated with 12.5 nM miR-622 or a negative control mimic. After 24 h, those cells were co-transfected with 0.2 μg of firefly luciferase reporter vector containing the HULC sequence or the empty vector and transfected with 0.1 μg of Renilla luciferase reporter pRL-SV40 (Promega) using Lipofectamine 2000 (Thermo Fisher Scientific). After another 24 h, luciferase activity was evaluated using the Dual-Luciferase Reporter Assay (Promega). Relative firefly luciferase activity was normalized to Renilla luciferase activity.
Takahashi K., Koyama K., Ota Y., Iwamoto H., Yamakita K., Fujii S, & Kitano Y. (2020). The Interaction Between Long Non-coding RNA HULC and MicroRNA-622 via Transfer by Extracellular Vesicles Regulates Cell Invasion and Migration in Human Pancreatic Cancer. Frontiers in Oncology, 10, 1013.
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10

Snail 3'UTR Luciferase Reporter Assay

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The Snail 3′untranslated region (3′UTR) firefly luciferase reporter vector and empty vector were purchased from OriGene Technologies (Rockville, MD, USA). Cells (2 × 105 cells·well−1) were seeded on a six‐well plate and treated with 12.5 nm of mirVana® miR‐195 or the negative control mimic. After 24 h, cells were cotransfected with 2.0 μg of Snail 3′UTR firefly luciferase reporter vector or empty vector, and 0.1 μg of Renilla luciferase reporter pRL‐SV40 (Promega, Madison, WI, USA) using Lipofectamine 2000 (Thermo Fisher Scientific). After another 24 h, luciferase activity was measured using the Dual‐Luciferase Reporter Assay (Promega). Relative firefly luciferase activity was normalized to Renilla luciferase activity.
Ota Y., Takahashi K., Otake S., Tamaki Y., Okada M., Yan I., Aso K., Fujii S., Patel T, & Haneda M. (2021). Extracellular RNA transfer from non‐malignant human cholangiocytes can promote cholangiocarcinoma growth. FEBS Open Bio, 11(12), 3276-3292.
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