Scc142

Manufactured by Merck Group

Sourced in United States

The SCC142 is a laboratory instrument designed for the detection and analysis of various chemical compounds. It features advanced spectroscopy technology to provide accurate measurements and data analysis. The core function of the SCC142 is to serve as a versatile analytical tool for researchers and scientists in various fields of study.

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3 protocols using scc142

Promoter Reporter Assays in HEK293FT and DC2.4 Cells

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HEK293FT or DC2.4 (SCC142, Millipore) cells were cultured in DMEM supplied with 10% FBS (GIBCO) and penicillin–streptomycin (Thermo Fisher Scientific). VCAM1, Il1b, Tnf, Il23, Ppara, Rara, IFNG, RORC and IL12RB promoter reporter plasmids expressing gaussia luciferase were acquired from GeneCopoeia; pGud-Luc has been described previously^{91 (link),92 (link)}. Reporters were transfected with Lipofectamine 2000 (11668019, Thermo Fisher Scientific). To evaluate AHR promoter activity, cells were stimulated with vehicle, propyzamide (10 μM), FICZ (0.5 μM) or a combination of propyzamide and FICZ. To evaluate VCAM1 promoter activity, cells were stimulated with vehicle or propyzamide for 2 more days before medium collection. CEBPB plasmid (15738, Addgene) or pGL4.54[luc2/TK] vector (E506A, Promega) were co-transfected to evaluate C/EBPβ binding to the Il1b, Tnf, Il23, RORC, IL12RB1 or IFNG promoters. Luciferase activity was evaluated using the Gaussia Luciferase Flash Assay Kit (16159, Thermo Fisher Scientific).

Sanmarco L.M., Chao C.C., Wang Y.C., Kenison J.E., Li Z., Rone J.M., Rejano-Gordillo C.M., Polonio C.M., Gutierrez-Vazquez C., Piester G., Plasencia A., Li L., Giovannoni F., Lee H.G., Akl C.F., Wheeler M.A., Mascanfroni I., Jaronen M., Alsuwailm M., Hewson P., Yeste A., Andersen B.M., Franks D.G., Huang C.J., Ekwudo M., Tjon E.C., Rothhammer V., Takenaka M., de Lima K.A., Linnerbauer M., Guo L., Covacu R., Queva H., Fonseca-Castro P.H., Bladi M.A., Cox L.M., Hodgetts K.J., Hahn M.E., Mildner A., Korzenik J., Hauser R., Snapper S.B, & Quintana F.J. (2022). Identification of environmental factors that promote intestinal inflammation. Nature, 611(7937), 801-809.

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Hypoxia-Inducible Factor 1-Alpha Regulation

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A total of 4 × 10⁶ BMDCs from FVB.129S6-Gt(ROSA)26Sortm2(HIF1A/luc)Kael/J mice (006206, The Jackson Laboratory) were stimulated with LPS or LPS + L-LA (10 mM) for 6 h. Next, the luciferase activity was quantified using the Dual-Luciferase Reporter Assay System (Promega, E1910) according to the manufacturer’s protocol.
In another set of experiments, HepG2 (ATCC HB-8065) or DC2.4 (SCC142, Millipore) cells were cultured in DMEM supplied with 10% FBS (Gibco) and penicillin–streptomycin (Thermo Fisher Scientific). Ndufa4l2- and Hif1a-promoter reporter plasmids expressing Gaussia luciferase were acquired from GeneCopoeia. To evaluate Ndufa4l2 promoter activity cells were stimulated with vehicle, L-LA (10 mM), LPS (100 ng ml⁻¹) or the combination of L-LA and LPS. For overexpression assays, HIF1A plasmid (OHu27176, GenScript), Ndufa4l2 plasmid (EX-J0135-M02, GeneCopoeia) or empty control vector (GeneCopoeia) were used. All plasmids were transfected using lipofectamine 2000 (11668019, Thermo Fisher Scientific) following the manufacturer’s instructions. Luciferase activity was evaluated with the Gaussia Luciferase Flash Assay Kit (16159, Thermo Fisher Scientific).

Zhu Y., Traub L.M, & Kornfeld S. (1999). High-Affinity Binding Of The AP-1 Adaptor Complex to Trans-Golgi Network Membranes Devoid Of Mannose 6-Phosphate Receptors. Molecular Biology of the Cell, 10(3), 537-549.

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Culturing and Maintaining Cell Lines

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The human HEK293T (CRL-3216), HEK293T/17 (CRL-11268), HeLa (CCL-2) obtained from ATCC and mouse DC2.4 (SCC142, Millipore, Burlington, MA, USA) cell lines maintained at 37 °C in a humidified atmosphere with 5% CO₂. HEK293T and HeLa cells were cultured in DMEM (Gibco, Waltham, MA, USA) containing alanyl-glutamine (Paneco, Moscow, Russia) and supplemented with 10% FBS (HyClone, Logan, UT, USA) along with penicillin and streptomycin (Paneco, Moscow, Russia). DC2.4 cells were cultured in RPMI (Gibco, USA) medium with the same supplements. HEK293T/17 cells were cultured in DMEM/F12 (1:1) supplemented with 10% FBS, sodium pyruvate, Glutamax, antibiotic/antimycotic solution, and non-essential amino acids (all reagents from Gibco, USA). To prevent mycoplasma contamination, all cell cultures were constantly tested using the Myco-visor kit (Biolabmix, Novosibirsk, Russia).

Fedorovskiy A.G., Antropov D.N., Dome A.S., Puchkov P.A., Makarova D.M., Konopleva M.V., Matveeva A.M., Panova E.A., Shmendel E.V., Maslov M.A., Dmitriev S.E., Stepanov G.A, & Markov O.V. (2024). Novel Efficient Lipid-Based Delivery Systems Enable a Delayed Uptake and Sustained Expression of mRNA in Human Cells and Mouse Tissues. Pharmaceutics, 16(5), 684.

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