4xm67 ptata tk luc

Manufactured by Addgene

The 4xM67 pTATA TK-Luc is a plasmid that contains a luciferase reporter gene under the control of a minimal promoter with four copies of the M67 binding site. This plasmid can be used to study the activity of transcription factors that bind to the M67 sequence.

Automatically generated - may contain errors

Lab products found in correlation

Stat3 c flag prc cmv, by Addgene (1 mentions) Prl sv40, by Addgene (1 mentions) Pgl4.29 luc2p cre hygro, by Promega (1 mentions) Pgl4.33 luc2p sre hygro, by Promega (1 mentions) Reporter lysis 5x buffer, by Promega (1 mentions) Fugene hd transfection reagent, by Promega (1 mentions) Luciferase assay reagent, by Promega (1 mentions) Transfection reagent, by Horizon Discovery (1 mentions) On targetplus smartpool sirna, by Thermo Fisher Scientific (1 mentions) Egm 2 bulletkit, by Lonza (1 mentions) Stattic, by Merck Group (1 mentions)

Spelling variants of this product

PTATA TK-Luc (4 citations)

4 protocols using 4xm67 ptata tk luc

Twist1 Promoter and Expression Constructs

Check if the same lab product or an alternative is used in the 5 most similar protocols

We generated Twist1 promoter reporter construct Twist1-Luc by cloning human Twist1 promoter −451 to +1 fragment into pGL3-basic luciferase vector at KpnI/XhoI sites. The full length Twist1 coding domain sequence was PCR amplified from pWZL Blast Twist1 ER construct (Addgene, USA) and inserted into pcDNA3.1 vector at BamHI/ECORI sites, yielding pcDNA3.1-Twist1. The STAT3 reporter construct 4xM67 pTATA TK-Luc and the constitutive activated STAT3 expression construct STAT3-C Flag pRc/CMV were obtained from Addgene.

Cao H.H., Chu J.H., Kwan H.Y., Su T., Yu H., Cheng C.Y., Fu X.Q., Guo H., Li T., Tse A.K., Chou G.X., Mo H.B, & Yu Z.L. (2016). Inhibition of the STAT3 signaling pathway contributes to apigenin-mediated anti-metastatic effect in melanoma. Scientific Reports, 6, 21731.

+ Open protocol

+ Expand

Plasmid Transfection Assay Protocol

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

CISH reporter plasmid was kindly supplied by Dr. Clevenger [21 (link)]. pGL4.33[luc2P/SRE/Hygro] (MEK-Erk response element, Promega #E1340); pGL4.29[luc2P/CRE/Hygro] (cAMP response element, Promega #E8471); 4xM67 pTATA TK-Luc (Stat3 response element, Addgene #8688) are commercially available. pRL-SV40 (Addgene #E2231) was used as transfection efficiency control.

Cuesta-Casanovas L., Delgado-Martínez J., Cornet-Masana J.M., Carbó J.M., Banús-Mulet A., Guijarro F., Esteve J, & Risueño R.M. (2023). Prolactin receptor signaling induces acquisition of chemoresistance and reduces clonogenicity in acute myeloid leukemia. Cancer Cell International, 23, 97.

+ Open protocol

+ Expand

HEK293T Cell Signaling Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

HEK293T cells were maintained at 37°C and 5% CO₂ in DMEM, containing 10% FBS (HyClone) and 100 mg ml-1 penicillin-streptomycin. In general, siRNA was transfected using transfection reagent (Dharmacon, T-2001-02) following manufacturer’s protocol, and asprosin was overexpressed using Ad5-Asprosin or Asprosin coding mammalian expression plasmid with Ad5-empty or CMV6-Entry-empty as controls. For luciferase readout, HEK293T cells were transduced with 4xM67 pTATA-TK-Luc (Addgene; 8688) plasmid. HEK293T cell transfection, lysis and luciferase readouts were done with Fugene HD transfection reagent (Promega; E2312), Reporter Lysis 5X Buffer (Promega, E3971) and Luciferase Assay Reagent (Promega, E1483), respectively, using standard manufacturer’s protocol.

Mishra I., Xie W.R., Bournat J.C., He Y., Wang C., Silva E.S., Liu H., Ku Z., Chen Y., Erokwu B.O., Jia P., Zhao Z., An Z., Flask C.A., He Y., Xu Y, & Chopra A.R. (2022). Protein Tyrosine Phosphatase Receptor δ Serves as the Orexigenic Asprosin Receptor. Cell metabolism, 34(4), 549-563.e8.

+ Open protocol

+ Expand

Studying KSHV Infection Pathways

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human umbilical vein endothelial cells (HUVECs; Lonza, Walkersville, MD) were maintained for five passages in complete EGM-2 BulletKit (Lonza). The KSHV-latently infected cell line, BCBL-1, was maintained in RPMI 1640 supplemented with 10% FBS, 1X Peniciliin-Streptomycin and 55 μM β-mercaptoethanol. 293 cells were maintained in DMEM supplemented with 10% FBS and 1X Penicillin-Streptomycin. miRVana miRNA mimics of various KSHV-encoded miRNAs were purchased from Ambion (Life Technologies Inc., Carlsbad, CA). ON-TARGETplus SMARTpool siRNAs targeting human STAT3, BIRC5, IRAK1 and the non-targeting pool control were obtained from Thermo Fisher Scientific (Waltham, MA). The STAT3 inhibitor- Stattic, was purchased from Sigma-Aldrich Inc, St. Louis, MO. The STAT3-dependent luciferase expression plasmid, 4xM67 pTATA TK-Luc, was a gift from Jim Darnell and was obtained through Addgene.

Ramalingam D, & Ziegelbauer J.M. (2017). Viral microRNAs Target a Gene Network, Inhibit STAT Activation, and Suppress Interferon Responses. Scientific Reports, 7, 40813.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!