P9389

Manufactured by Merck Group

P9389 is a lab equipment product from Merck Group. It is designed for general laboratory use. The core function of this product is to perform standard laboratory tasks, but its specific intended use is not provided in this unbiased and factual description.

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Lab products found in correlation

Curdlan, by Fujifilm (1 mentions) Pmaxgfp vector, by Lonza (1 mentions) Poly da dt, by Merck Group (1 mentions) Recombinant murine gm csf, by Thermo Fisher Scientific (1 mentions) Cell culture reagents, by Thermo Fisher Scientific (1 mentions) D1626, by Merck Group (1 mentions)

3 protocols using p9389

Poxviral DNA Isolation and Purification

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All reagents, including Poly(dG-dC) · Poly(dG-dC) acid sodium salt (poly(dG:dC), P9389), calf thymus DNA (CT-DNA, D4764), Poly(dA-dT) · Poly(dA-dT) acid sodium salt (poly(dA:dT), P0883) were purchased from Sigma, if not otherwise stated. cGAMP was obtained from Invivogen (tlrl-cga-s).The pmaxGFP Vector from Lonza was used as the circular plasmid DNA. Cell culture reagents were obtained from Invitrogen, and FCS was obtained from HyClone. Murine recombinant GM-CSF was purchased from PreproTech. CpG 1826 oligonucleotides, ultrapure LPS, and endotoxin-free DNA from E. coli K12 were purchased from InvivoGen, and curdlan was purchased from Wako. Poxviral genomic DNA from VV strain CVA and from cowpox virus (isolate 81/01, 5^th passage in MA104 cells; originally provided by S. Essbauer, Munich, Germany) was isolated and purified from infected cell cultures, as recently described^{48 (link)}.

Roth S., Rottach A., Lotz-Havla A.S., Laux V., Muschaweckh A., Gersting S.W., Muntau A.C., Hopfner K.P., Jin L., Vanness K., Petrini J.H., Drexler I., Leonhardt H, & Ruland J. (2014). Rad50-CARD9 interactions link cytosolic DNA sensing to IL-1β production. Nature immunology, 15(6), 538-545.

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DNA-Cu2TPNap Complex Interactions

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Complex-DNA interactions were analysed in 10 mM phosphate solution (pH 7.0) in the presence of 25 mM NaCl. Solutions of salmon testes DNA (stDNA, Sigma Aldrich, D1626, ϵ₂₆₀ = 12 824 M(bp)⁻¹ cm⁻¹), Poly[(d(A-T)₂] (Sigma Aldrich, PO883, ϵ₂₆₀ = 13 100 M(bp)⁻¹ cm⁻¹) and Poly[(d(G-C)₂] (Sigma Aldrich, P9389, ϵ₂₆₀ = 16 800 M(bp)⁻¹ cm⁻¹) were initially heat treated and allowed to renature prior to quantification using an Agilent Cary 100 dual beam spectrophotometer equipped with a 6 × 6 peltier multicell system with temperature controller to give a working solution with final DNA concentration of ∼100 μM (bp equivalents). The investigation was conducted in the range of 200–400 nm and measurements were recorded at a rate of 1 nm per second. DNA solutions were incubated for 30 min periods in darkness at 37°C with Cu₂TPNap at varing r ([drug] / [DNA]) values where r = 0.1 and 0.2 (r being the ratio of complex per μM DNA).

Molphy Z., Montagner D., Bhat S.S., Slator C., Long C., Erxleben A, & Kellett A. (2018). A phosphate-targeted dinuclear Cu(II) complex combining major groove binding and oxidative DNA cleavage. Nucleic Acids Research, 46(19), 9918-9931.

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Fluorescence-based Nucleic Acid Binding Assay

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This assay was carried out according to the procedure reported by Molphy et al. [41] Briefly, a working solution was prepared containing 100 µM poly[d(G-C)2] (Sigma Aldrich P9389, ε254 = 8,400 M -1 cm -1 ) in 10 mM PO4 3-, 50 mM NaCl and 10 mM MgCl2 (pH 7.4). Stocks of mithramycin A (Sigma Aldrich, M6891, ε400 = 10,000 M -1 cm -1 ) and of the complexes were prepared in DMF. Cu-DPA-Phenazine derivatives were added to the working solution at a ratio of 0.10 drug/nucleotide. Varying amounts of mithramycin A (r = 0-0.25) were added to complex treated and complex untreated poly[d(G-C)2] and allowed to incubate in the dark for 5 minutes prior to analysis. Emission spectra were recorded in the range 525 -625 nm with an excitation wavelength of 470 nm to avoid photodegradation (excitation slit: 2.5 nm; emission slit: 5 nm). Triplicate values were recorded and all data was analysed at 550 nm.

Fantoni N.Z., Molphy Z., O'Carroll S., Menounou G., Mitrikas G., Krokidis M.G., Chatgilialoglu C., Colleran J., Banasiak A., Clynes M., Roche S., Roche S., Kelly S., McKee V, & Kellett A. (2021). Polypyridyl-Based Copper Phenanthrene Complexes: Combining Stability with Enhanced DNA Recognition. Chemistry (Weinheim an der Bergstrasse, Germany), 27(3).

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