Pcbfre luc

Manufactured by Addgene

Sourced in United States

The PCBFRE-luc is a plasmid designed for the expression and measurement of firefly luciferase activity. It contains a luciferase reporter gene under the control of a constitutive promoter, allowing for the quantification of gene expression in a variety of cell types and applications.

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Lab products found in correlation

Mrx revelation, by Dynex (1 mentions) Lumat3 lb9508 luminometer, by Berthold Technologies (1 mentions) Myrakt delta4 129, by Addgene (1 mentions) Wortmannin, by Merck Group (1 mentions) Pcdna3 flag mtor wt, by Addgene (1 mentions) Notch1, by Cell Signaling Technology (1 mentions) P p70s6k, by Cell Signaling Technology (1 mentions) N n 3 5 difluorophenacetyl l alanyl s phenylglycine t butyl ester dapt, by Merck Group (1 mentions) Odd luciferase pcdna3, by Addgene (1 mentions) P 4e bp1, by Santa Cruz Biotechnology (1 mentions) 3xflagnicd1, by Addgene (1 mentions) Pmxs hc myc, by Addgene (1 mentions) Ha hif1alpha pcdna3, by Addgene (1 mentions) 4e bp1, by Cell Signaling Technology (1 mentions) C myc, by Santa Cruz Biotechnology (1 mentions) Hif 1α, by Cell Signaling Technology (1 mentions) Dual luciferase reporter assay system, by Promega (1 mentions)

3 protocols using pcbfre luc

Luciferase Assay for CSL-Driven Transcription

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Luciferase reporter vector with a promoter containing four CSL-binding elements was purchased from Addgene (Cambridge, MA, USA) (pCBFRE-luc, plasmid#26897). The luciferase assay was conducted 36 h posttransient transfection. Lysis buffer was purchased from Promega (Madison, WI, USA) (E3971), and 20 of 150 μL of the lysis supernatant were used with a Lumat3 LB9508 luminometer (Berthold, Oak Ridge, TN, USA) for the measurement of luciferase activity. Each sample was measured three times. All results were normalized to β-galactosidase activity (MRX Revelation; DYNEX Technologies, CHANTILLY, VA, USA).

Liu B., Lin X., Yang X., Dong H., Yue X., Andrade K.C., Guo Z., Yang J., Wu L., Zhu X., Zhang S., Tian D., Wang J., Cai Q., Chen Q., Mao S., Chen Q, & Chang J. (2015). Downregulation of RND3/RhoE in glioblastoma patients promotes tumorigenesis through augmentation of notch transcriptional complex activity. Cancer Medicine, 4(9), 1404-1416.

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Isolation and Purification of TF3 Monomer

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TF3 monomer (purity: 92.4%) was isolated and purified using previously established method (15 (link)). Wortmannin and N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (DAPT) were purchased from Sigma and Santa Cruz Biotechnology (Santa Cruz, CA, USA), respectively. Antibodies against phospho-Akt (Ser473) (p-Akt), Akt, phospho-p70S6 kinase (Thr421/Ser424) (p-p70S6K), p70S6K, eukaryotic initiation factor 4E-binding protein-1 (4E-BP1), HIF-1α, Notch-1, c-Jun N-terminal kinases (JNK), p38 and phosphor-Forkhead Box O1 (Thr24) (p-FoxO1) were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). Antibodies against phosphor-mammalian target of rapamycin (p-mTOR) (Ser2448), mTOR were purchased from R&D Systems (Minneapolis, MN, USA). Antibodies against phosphor-4E-BP1 (Ser65/Thr70) (p-4E-BP1), c-Myc, phosphor-extracellular signal-regulated kinases 1/2 (Thr202/Tyr204) (p-ERK1/2), ERK1/2 and GAPDH were purchased from Santa Cruz Biotechnology Inc. (Santa Cruz). Plasmids (myrAkt delta4-129, pcDNA3-Flag mTOR wt, pWZL Neo Myr Flag RPS6KB1, pET14b PHAS-I, HA-HIF1alpha-pcDNA3, 3XFlagNICD1, pMXs-hc-Myc, ODD-Luciferase-pcDNA3, VEGF promoter, cMyc promoter (TBE1/2-wt), and pCBFRE-luc) were purchased from Addgene (Cambridge, MA, USA).

GAO Y., RANKIN G.O., TU Y, & CHEN Y.C. (2015). Theaflavin-3, 3′-digallate decreases human ovarian carcinoma OVCAR-3 cell-induced angiogenesis via Akt and Notch-1 pathways, not via MAPK pathways. International Journal of Oncology, 48(1), 281-292.

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Notch Transcriptional Activity Assay

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To assess the effect of BLVRB on Notch-mediated transcriptional activity, we used a traditional dual-luciferase assay consisting of four Notch-sensing CBF1-binding site reporters (normalized to a control promoter driving Renilla luciferase), as previously described 27 (link). pCBFRE-luc was donated by Nicholas Gaiano (plasmid# 26897; Addgene, Watertown, MA, USA). Briefly, cells were co-transfected with the CBF1-luciferase reporter construct and a Renilla luciferase reporter plasmid. Luciferase activity was examined 24 h after transfection using a dual-luciferase reporter assay system (Promega, Madison, WI, USA) according to the manufacturer's instructions. Firefly luciferase activity from the CBF1 reporter construct was normalized to the control Renilla luciferase included in the kit. Luciferase activity was expressed as the fold increase relative to unstimulated conditions.

Gao Z., Ni X., Zheng B., Sun W., Wan W., Liu H., Ni X., Suo T., Li N., Liu H, & Shen S. (2022). Biliverdin reductase B impairs cholangiocarcinoma cell motility by inhibiting the Notch/Snail signaling pathway. Journal of Cancer, 13(7), 2159-2170.

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