Victor x machine

Manufactured by PerkinElmer

Sourced in United States

The Victor X is a high-performance multi-mode microplate reader designed for life science research applications. It offers versatile detection capabilities, including absorbance, fluorescence, and luminescence measurements, enabling a wide range of assays to be performed. The Victor X provides accurate and reliable data, making it a valuable tool for researchers in various fields.

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Lab products found in correlation

Pgl3 vector, by Genechem (4 mentions) Lipofectamine ltx, by Thermo Fisher Scientific (3 mentions) Dual luciferase reporter assay system, by Promega (2 mentions) Dual luciferase reporter assay system kit, by Promega (2 mentions) Pgl3 vector, by Promega (1 mentions) Quickchange xl site directed mutagenesis kit, by Agilent Technologies (1 mentions) Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions) Dual luciferase reporter system, by Promega (1 mentions) Dual luciferase reporter assay kit, by Promega (1 mentions)

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Victor X

6 protocols using victor x machine

Characterizing SPRED1 3'UTR Regulation

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The wild-type or mutant 3′-untranslated region (UTR) sequences of SPRED1 were
inserted into the restriction sites located at the 3′ end of the luciferase gene
of the pGL3 vector (GeneChem). The wild-type or mutant sequence is shown in
Fig. 4B. The pGL3
vector with the wild-type or mutant 3′-UTR sequences of SPRED1 and pRL-TK
vectors were co-transfected into the corresponding cells using Lipofectamine LTX
(Invitrogen). Luciferase activity was assayed 48 h after transfection according
to the manufacturer’s protocol (Promega, Madison, WI, USA). Firefly and Renilla
luciferase activities were detected using a Dual-luciferase Reporter Assay
System Kit (Promega) with a Victor X machine (Perkin-Elmer, Boston, MA,
USA).

Zhu X., Peng C., Peng Z., Chang R, & Guo Q. (2022). Sevoflurane Inhibits Metastasis in Hepatocellular Carcinoma by Inhibiting MiR-665-Induced Activation of the ERK/MMP Pathway. Cell Transplantation, 31, 09636897221104447.

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Analyzing miR-204-5p Regulation of SIX1 Activity

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The effects of miR‐204‐5p overexpression and knockdown on the SIX1 activity were measured by dual‐luciferase reporter assays. The BEL7402 cells were transfected with the wild‐type (WT) or MUT 3′ UTR of SIX1, which was inserted into the pGL3 vector in 96‐well plates. After 24 h, the dual‐luciferase assays were performed using the dual‐luciferase reporter assay system (Promega, Madison, WI,USA) with a Victor X machine (PerkinElmer, Norwalk, CT, USA).

Chu Y., Jiang M., Du F., Chen D., Ye T., Xu B., Li X., Wang W., Qiu Z., Liu H., Nie Y., Liang J, & Fan D. (2018). miR‐204‐5p suppresses hepatocellular cancer proliferation by regulating homeoprotein SIX1 expression. FEBS Open Bio, 8(2), 189-200.

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miR-204-5p Regulates CXCR4 and CXCL12

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The effects of miR-204-5p overexpression on the CXCR4 and CXCL12 activity were measured by dual-luciferase reporter assays. The plasmids of the wild-type (3'UTR-W) and corresponding mutant-type (3'UTR-M) containing CXCR4 and CXCL12 3'UTR cDNA were constructed based on the pGL3 vector (Promega Corp., Madison, WI, USA; GenBank® Accession Number U47296, Catalog number selected: E1741). The 293T cells were con-transfected with the miR-204-5p mimics or negative control (Nc) and the plasmid 3'UTR-W and 3'UTR-M of CXCR4 or CXCL12. After 24 h, the dual-luciferase assays were performed using the dualluciferase reporter assay system (Promega, Madison, WI, USA) with a Victor X machine (PerkinElmer, Norwalk, CT, USA).

Zhang J., Xing L., Xu H., Wang K., She J., Shi F., Wu H., Sun Y., Gao J, & He S. (2020). miR-204-5p Suppress Lymph Node Metastasis via Regulating CXCL12 and CXCR4 in Gastric Cancer. Journal of Cancer, 11(11), 3199-3206.

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Identifying miR-144 target ADAM10

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Three online prediction programs, TargetScan, miRanda, and PicTar, were used to identify candidate targets for miR-144. The gene for ADAM10, predicted by all three databases and associated with cognitive function, was further studied as a potential target. Sequence of segments with WT or mutant 3’-UTR region of ADAM10 were synthesized and cloned into the pGL3 vector (GeneChem, Shanghai, China). All constructs were verified by sequencing. First, the N2A or HEK293 cells were seeded at 0.5×10⁵ cell per well in 24-well plates 24 h prior to transfection. The following day, the pGL3 vector containing WT 3’-UTR of ADAM10 mRNA or mutant forms was co-transfected with LV-miR-144 into N2A or HEK293 cells. After 48 h, all cells were harvested according to manufacturer's protocol (Promega, Madison, WI), and the Firefly and Renilla luciferase activity were determined using dul-luciferase reporter assay system (Promega, Madison, WI) with a Victor X machine (PerkinElmer, Boston, MA). Firefly luciferase activity was normalized to Renilla luciferase activity. Three independent experiments were performed in triplicate.

Sun L., Zhao M., Zhang J., Liu A., Ji W., Li Y., Yang X, & Wu Z. (2017). MiR-144 promotes β-amyloid accumulation-induced cognitive impairments by targeting ADAM10 following traumatic brain injury. Oncotarget, 8(35), 59181-59203.

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Validating SND1 as miR-361-5p Target

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Dual-luciferase 3′ UTR reporter assay was carried out to validate SND1 as a direct target of miR-361-5p. Wild-type (WT) or mutant of 3′ untranslated region (UTR) sequences of SND1 were inserted into the Fse I and Xba I sites of the pGL3 vector (GeneChem). HEK293 cells infected with anti-miR-361-5p lentivirus or negative control (NC) lentivirus were plated into 96-well plates. pGL3 vector (40 ng) with the above sequence was cotransfected with 10 ng of pRL-TK vector into cells by Lipofectamine LTX (Invitrogen). Twenty-four hours later, cells were collected according to the manufacturer's protocol (Promega) and firefly and Renilla luciferase activity was detected using Dual-luciferase Reporter Assay System Kits (Promega) with a Victor X machine (PerkinElmer). Additionally, a mutant SND1 3′ UTR reporter construct was made by site-directed mutagenesis in the putative target site of miR-361-5p using Quickchange XL site-directed mutagenesis kit (Agilent Technologies). All PCR products were verified by DNA sequencing. The normalized luciferase activity was expressed as a ratio of firefly luciferase to Renilla luciferase units.

Ma F., Song H., Guo B., Zhang Y., Zheng Y., Lin C., Wu Y., Guan G., Sha R., Zhou Q., Wang D., Zhou X., Li J, & Qiu X. (2015). MiR-361-5p inhibits colorectal and gastric cancer growth and metastasis by targeting staphylococcal nuclease domain containing-1. Oncotarget, 6(19), 17404-17416.

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Luciferase Assay for miRNA and NF-κB Activity

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The wild-type or mutant 3′-UTR sequences of SPHK1 were ligated into the XbaI and FseI sites of the pGL3 vector (GeneChem). HEK293T cells infected with the miR-506 lentivirus or the negative control lentivirus were seeded into 96-well plates. The cells were co-transfected with 50 ng of the pGL3 vector and 10 ng of the pRL-TK vector using Lipofectamine LTX (Invitrogen). Twenty-four hours after transfection, the cells were harvested according to the manufacturer's protocol (Promega, Madison, WI, USA), and firefly and Renilla luciferase activities were measured using a Dual Luciferase Reporter System (Promega) with a Victor X machine (Perkin-Elmer, Boston, MA, USA). For the NF-κB transcriptional activity assay, the pGL4.32[luc2P/NF-κB-RE/Hygro] plasmid or control luciferase plasmid pGL4.74[hRluc/TK] (Promega) was transfected using the Lipofectamine 2000 reagent (Invitrogen), and luciferase and Renilla signals were measured using a Dual Luciferase Reporter Assay kit (Promega) according to the manufacturer's protocol.

Li J., Wu H., Li W., Yin L., Guo S., Xu X., Ouyang Y., Zhao Z., Liu S., Tian Y., Tian Z., Ju J., Ni B, & Wang H. (2016). Downregulated miR-506 expression facilitates pancreatic cancer progression and chemoresistance via SPHK1/Akt/NF-κB signaling. Oncogene, 35(42), 5501-5514.

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