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Ptk renilla

Manufactured by Promega
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The PTK-Renilla is a lab equipment product manufactured by Promega. It is a non-radioactive, bioluminescent reporter assay system used to measure protein tyrosine kinase (PTK) activity in cells.

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Lab products found in correlation

Dual luciferase reporter assay kit, by Promega (5 mentions) Dual luciferase reporter assay, by Promega (4 mentions) X tremegene hp dna transfection reagent, by Roche (2 mentions) Spectramax l microplate reader, by Molecular Devices (2 mentions) Dual luciferase reporter system, by Promega (2 mentions) Fugene transfection reagent, by Promega (2 mentions) Lipofectamine ltx reagent, by Thermo Fisher Scientific (2 mentions) Dual glo luciferase assay system, by Promega (2 mentions) X tremegene 9 dna transfection reagent, by Roche (2 mentions) Scrambled sirna sc 37007, by Santa Cruz Biotechnology (1 mentions) Pgl3 basic vector, by Promega (1 mentions) Pdonr207, by Thermo Fisher Scientific (1 mentions) T7 ribomax express large scale rna production system, by Promega (1 mentions) Recombinant tnf α, by R&D Systems (1 mentions) Recombinant ifn β, by PBL Assay Science (1 mentions) Verikine human ifn β elisa kit, by PBL Assay Science (1 mentions) 17β estradiol e2, by Merck Group (1 mentions) Rpmi 1640, by Merck Group (1 mentions) Tamoxifen tam, by Merck Group (1 mentions) Charcoal stripped fbs, by Sartorius (1 mentions)

Close spelling variants of this product

PTK Renilla luciferase plasmid (2 citations) PTK-Renilla-Luc (2 citations) PTK-Renilla normalization plasmid (2 citations)

21 protocols using ptk renilla

1

Transient Transfection and Luciferase Assay

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HeLa cells were seeded in duplicate in 24‐well tissue culture plates, followed by transient transfection with hZEB1‐Luc, pTKRenilla (Promega, San Luis Obispo, CA, USA), and the indicated expression plasmids using X‐treme Gene HP DNA transfection reagent (Roche, Indianapolis, IN, USA). Twenty‐four hours later, luciferase activity was determined using the Dual Luciferase Reporter Assay (Promega) on a luminometer (SpectraMax L Microplate Reader; Molecular Devices, Sunnyvale, CA, USA). Firefly luciferase activity was normalized to sea‐pansy luciferase activity from cotransfected pTKRenilla.
Sinh N.D., Endo K., Miyazawa K, & Saitoh M. (2017). Ets1 and ESE1 reciprocally regulate expression of ZEB1/ZEB2, dependent on ERK1/2 activity, in breast cancer cells. Cancer Science, 108(5), 952-960.
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2

Dual Luciferase Reporter Assay Protocol

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For Dual luciferase reporter assays, ~80000 cells plated in each well of 24-well plates and transfected with 200ng reporter plasmid, 20ng normalization plasmid (pTK-Renilla; Promega) and 200ng expression plasmid. Dual luciferase reporter assays (Promega) were performed 48h after transfection according to manufacturer’s instructions. For each well, firefly (reporter) luciferase activity was divided by renilla (normalizer) luciferase activity to determine the normalized level of promoter activation. Each condition in the reporter assays had six biologic replicates.
Grassmeyer J., Mukherjee M., deRiso J., Hettinger C., Bailey M., Sinha S., Visvader J.E., Zhao H., Fogarty E, & Surendran K. (2017). Elf5 is a principal cell lineage specific transcription factor in the kidney that contributes to Aqp2 and Avpr2 gene expression. Developmental biology, 424(1), 77-89.
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3

Dual-Luciferase Assay for AhR Activity

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Fifteen thousand human embryonic kidney (HEK)-293 cells per well were plated in 96-well plates (flat bottom). Twenty-four hours after plating, cells were transfected with equal amounts of pGud-Luc (Firefly luciferase under control of AHR-responsive promoter element27 (link)) and pTK-Renilla (Renilla luciferase under control of constitutively active thymidine kinase promoter; Promega, Madison, WI) using Fugene Transfection Reagent (Promega) as suggested by the manufacturer. After 24 hours, transfected cells were incubated with Dulbecco's modified eagle medium (DMEM) supplemented with 10% of patient serum in duplicates. Luciferase activity was analyzed 24 hours later using the Dual-Luciferase Reporter System (Promega). Firefly luciferase activity was divided by Renilla luciferase activity and normalized to their respective control levels, which were set as 100%.
The study was approved by the Institutional Review Board of Brigham and Women's Hospital, and all participants provided written informed consent.
Rothhammer V., Borucki D.M., Garcia Sanchez M.I., Mazzola M.A., Hemond C.C., Regev K., Paul A., Kivisäkk P., Bakshi R., Izquierdo G., Weiner H.L, & Quintana F.J. (2017). Dynamic regulation of serum aryl hydrocarbon receptor agonists in MS. Neurology® Neuroimmunology & Neuroinflammation, 4(4), e359.
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4

NF-κB Luciferase Reporter Assay

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The NF-κB luciferase reporter vector (cat. no. 219078) was obtained from Agilent Technologies, Inc. (Santa Clara, CA, USA). Cells were transfected with NF-κB reporter vector using Lipofectamine® LTX reagent (Invitrogen; Thermo Fisher Scientific, Inc.). GL3-basic vector (Promega Corporation, Madison, WI, USA) served as a negative control. Cells were treated with P. endodontalis LPS for 1 h. The efficiency of transfection was standardized by co-transfection with pTK-Renilla (Promega Corporation). Total cell lysates were prepared using the Dual-Glo® Luciferase assay system (Promega Corporation) and subsequently assessed for luciferase activity.
Ma N., Yang D., Okamura H., Teramachi J., Hasegawa T., Qiu L, & Haneji T. (2016). Involvement of interleukin-23 induced by Porphyromonas endodontalis lipopolysaccharide in osteoclastogenesis. Molecular Medicine Reports, 15(2), 559-566.
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5

Plasmid Constructs and siRNA Knockdown

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GFP-Rab7 expression construct was a gift from C. Arregui (Universidad de San Martin, Buenos Aires, Argentina; Hernández et al., 2006 (link)). Plasmid pTK-ERE-luc containing five copies of the ERE upstream of the luciferase cassette was a gift from C. Jordan (University of Texas, Houston, TX). pTK-renilla was purchased from Promega.The constructs were verified by sequencing. siRNA/Stealth against ERα was purchased from Invitrogen as the following sequences: sense 5′-CAGAGGCUCUCAAACUAUAAAGAAA-3′, and antisense 5′-UUUCUUUAUAGUUUGAGAGCCUCUG-3′. siRNA against caveolin 1 (sc-29241), siRNA against clathrin–heavy chain (HC; sc-35067), and scrambled siRNA (sc-37007) were purchased from Santa Cruz Biotechnology.
Sampayo R.G., Toscani A.M., Rubashkin M.G., Thi K., Masullo L.A., Violi I.L., Lakins J.N., Cáceres A., Hines W.C., Coluccio Leskow F., Stefani F.D., Chialvo D.R., Bissell M.J., Weaver V.M, & Simian M. (2018). Fibronectin rescues estrogen receptor α from lysosomal degradation in breast cancer cells. The Journal of Cell Biology, 217(8), 2777-2798.
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6

Regulation of IL-10 Gene Expression

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The DNA sequence containing wild-type (WT) and mutated RXRA::VDR binding element and its respective up- and down-stream sequence (300 bps in total) of human IL-10 gene promoter were synthesized, and sub-cloned into the pGL3-basic vector (Promega, Cat# E1751), and designated as pGL3-hu-IL10-WT and pGL3-hu-IL10-Mut, respectively. The resultant vectors were verified by DNA sequencing.
AC16 cells were co-transfected with respective gene promoter vectors and control or VDR gene-specific siRNAs, followed by various treatments as indicated in the figures. pTK-Renilla (20 ng per well; Promega, Cat# E2241) was included in all transfection assays as an internal control. Dual-luciferase activity assays were conducted 48 h after transfection using a standard protocol. The relative luciferase unit (RLU) was defined as the ratio of luciferase versus renilla activity to that of the control (set as 1.0).
Yang S., Wang C., Ruan C., Chen M., Cao R., Sheng L., Chang N., Xu T., Zhao P., Liu X., Zhu F., Xiao Q, & Gao S. (2022). Novel Insights into the Cardioprotective Effects of Calcitriol in Myocardial Infarction. Cells, 11(10), 1676.
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7

Plasmid-based Interferon Reporter Assay

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The pISRE-Luc and pNF-κB-Luc reporter plasmids were from Stratagene (Ref 219089 and 219078, respectively). Recombinant IFN-β was from PBL Assay Science (Ref 11410-2). Recombinant TNF-α was from R&D Systems (Ref 210-TA). Short synthetic 5′-triphosphate RNA molecules (ssRNA) were synthesized from pCI-neo vector digested with XbaI using T7 RiboMAX Express large scale RNA production system (Promega), and then purified with a filtering membrane (Millipore). Sheep polyclonal antibodies against IFN-α (31100-1) and IFN-β (31400-1) were from PBL Assay Science. The VeriKine Human IFN-β ELISA kit was from PBL Assay Science. DNA or RNA transfections were performed with JetPrime PEI following manufacturer’s recommendations (Polyplus transfection). The following plasmids were transfected to activate ISRE promoter sequences: pCiNeo-3xFlag-GW, pTK-Renilla (Promega), pDONR207 (ThermoFisher).
Khiar S., Lucas-Hourani M., Nisole S., Smith N., Helynck O., Bourgine M., Ruffié C., Herbeuval J.P., Munier-Lehmann H., Tangy F, & Vidalain P.O. (2017). Identification of a small molecule that primes the type I interferon response to cytosolic DNA. Scientific Reports, 7, 2561.
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8

Simultaneous Signaling Pathway Measurement

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To measure the activity of YAP1/WWTR1, Wnt/Ctnnb1, and NFAT signaling, cells seeded in 96-well plates were cotransfected with the luciferase reporters of 8XGTIIC, 8XSuper Top-Flash or pGL3-NFAT, respectively, with pTK-Renilla (Promega) and effector plasmids. Luciferase activity was measured with a dual-luciferase reporter assay kit (Promega) according to the manufacturer’s instructions.
Zhou T., Gao B., Fan Y., Liu Y., Feng S., Cong Q., Zhang X., Zhou Y., Yadav P.S., Lin J., Wu N., Zhao L., Huang D., Zhou S., Su P, & Yang Y. (2020). Piezo1/2 mediate mechanotransduction essential for bone formation through concerted activation of NFAT-YAP1-ß-catenin. eLife, 9, e52779.
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9

Measuring Wnt Signaling Activity

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Luciferase assays protocol was as described previously (Cha et al., 2011 (link)). To measure luciferase activity HEK293T cells were cotransfected with TOPFlash or FOPFlash luciferase reporter plasmids together with PROX1, β-catenin, AXIN or ΔN-TCF7L2 expression plasmids. X-tremeGene 9 DNA Transfection Reagent (Roche, Mannheim, Germany) was used according to manufacturer’s instruction for transfection. pTK-Renilla (Promega, Madison, WI, USA) as an internal control to determine transfection efficiency. Luciferase activity was measured 36 hr later using Dual luciferase reporter assay kit (Promega, Madison, WI, USA) according to the manufacturer’s instructions.
Cha B., Geng X., Mahamud M.R., Zhang J.Y., Chen L., Kim W., Jho E.H., Kim Y., Choi D., Dixon J.B., Chen H., Hong Y.K., Olson L., Kim T.H., Merrill B.J., Davis M.J, & Srinivasan R.S. (2018). Complementary Wnt Sources Regulate Lymphatic Vascular Development via PROX1-Dependent Wnt/β-Catenin Signaling. Cell reports, 25(3), 571-584.e5.
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10

Measuring Wnt Signaling Activity

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Luciferase assays protocol was as described previously (Cha et al., 2011 (link)). To measure luciferase activity HEK293T cells were cotransfected with TOPFlash or FOPFlash luciferase reporter plasmids together with PROX1, β-catenin, AXIN or ΔN-TCF7L2 expression plasmids. X-tremeGene 9 DNA Transfection Reagent (Roche, Mannheim, Germany) was used according to manufacturer’s instruction for transfection. pTK-Renilla (Promega, Madison, WI, USA) as an internal control to determine transfection efficiency. Luciferase activity was measured 36 hr later using Dual luciferase reporter assay kit (Promega, Madison, WI, USA) according to the manufacturer’s instructions.
Cha B., Geng X., Mahamud M.R., Zhang J.Y., Chen L., Kim W., Jho E.H., Kim Y., Choi D., Dixon J.B., Chen H., Hong Y.K., Olson L., Kim T.H., Merrill B.J., Davis M.J, & Srinivasan R.S. (2018). Complementary Wnt Sources Regulate Lymphatic Vascular Development via PROX1-Dependent Wnt/β-Catenin Signaling. Cell reports, 25(3), 571-584.e5.
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