Brassinin

Manufactured by Merck Group

Sourced in United States

Brassinin is a laboratory equipment product manufactured by Merck Group. It is a reagent used in biochemical and molecular biology research applications. Brassinin is a compound that can be utilized as a research tool, however, a detailed description of its core function is not available while maintaining an unbiased and purely factual approach.

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6 protocols using brassinin

Brassinin Antimicrobial Activity Assay

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To analyze brassinin antimicrobial activity against bacteria, the Pcal suspension was standardized to an OD₆₀₀ of 0.01 in LB and coincubated with or without 200 µM brassinin (Merck KGaA). After 24 h, bacterial growth was measured at OD₆₀₀.

Sakata N., Haraguchi T., Masuo S., Ishiga T, & Ishiga Y. (2022). Pseudomonas cannabina pv. alisalensis Virulence Factors Are Involved in Resistance to Plant-Derived Antimicrobials during Infection. Plants, 11(13), 1742.

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Antimicrobial Susceptibility of WT and NU19

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To analyze WT and NU19 susceptibility to plant-derived antimicrobials, WT and NU19 were grown at 28 °C on KB medium. The bacterial suspensions were standardized to an OD₆₀₀ of 0.01 with KB, and after 6 h incubation, 200 µM antimicrobials, including brassinin (Merck KGaA), sulforaphane (Funakoshi), camalexin (Merck KGaA), daidzein (INDOFINE Chemical Company, Hillsborough, NJ, USA), genistein (Tokyo Chemical Industry, Tokyo, Japan), indole (Tokyo Chemical Industry), and phloretin (Funakoshi), was added to each sample. Bacterial growth was measured at OD₆₀₀ after 24 h incubation.

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Brassinin Stock Solution Preparation

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Brassinin was purchased from Sigma–Aldrich (Sigma, St. Louis, MO, USA) and stock solution (100 mM) was prepared in sterile DMSO and stored at −80 °C until the experiments were conducted.

Kwon H.H., Ahn C.H., Lee H.J., Sim D.Y., Park J.E., Park S.Y., Kim B., Shim B.S, & Kim S.H. (2023). The Apoptotic and Anti-Warburg Effects of Brassinin in PC-3 Cells via Reactive Oxygen Species Production and the Inhibition of the c-Myc, SIRT1, and β-Catenin Signaling Axis. International Journal of Molecular Sciences, 24(18), 13912.

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Melanin Synthesis Pathway Assay

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Brassinin, 2,4,6-Tris(2-pyridyl)-s-triazine, Iron(II) sulfate heptahydrate, Iron(III) chloride hexahydrate, tyrosinase from mushroom, L-tyrosine, L-DOPA, phenylmethylsulfonyl fluoride, α-melanocyte stimulating hormone (α-MSH), and Triton X-100 were obtained from Sigma-Aldrich (St. Louis, MO, USA). All materials related to cell culture were purchased from Welgene (Daegu, Korea).

Lee M.K., Ryu H., Jeong H.H, & Lee B. (2022). Brassinin Abundant in Brassicaceae Suppresses Melanogenesis through Dual Mechanisms of Tyrosinase Inhibition. Foods, 12(1), 121.

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Fungal Growth and Phytoalexin Assay

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Botrytis cinerea strain AI18 (Kuroyanagi et al., 2022 (link)), Epichloë festucae strain Fl1 (Young et al., 2005 (link)) and their transformants used in this study are listed in Supplementary Table 1. They were grown on potato dextrose agar (PDA) at 23°C. For the incubation in phytoalexins, mycelia plugs (approx.1 mm³) were excised from the growing edge of the colony using a dissection microscope (Stemi DV4 Stereo Microscope, Carl Zeiss, Oberkochen, Germany) and submerged in 50 μl of water or indicated phytoalexin in a sealed 96 well clear plate. The plate was incubated at 23°C for the indicated time.
Capsidiol was purified from Nicotiana tabacum as previously reported (Matsukawa et al., 2013 (link)) and synthesized rishitin (Murai et al., 1975 (link)) was provided by former Prof. Akira Murai (Hokkaido University, Japan). Resveratrol and brassinin were obtained from Sigma-Aldrich (Burlington, MA, USA). Glyceollin (glyceollin I) was obtained from Wako pure chemical (Osaka, Japan). Medicarpin was obtained from MedChemExpress (Monmouth Junction, NJ, USA).

Bulasag A.S., Camagna M., Kuroyanagi T., Ashida A., Ito K., Tanaka A., Sato I., Chiba S., Ojika M, & Takemoto D. (2023). Botrytis cinerea tolerates phytoalexins produced by Solanaceae and Fabaceae plants through an efflux transporter BcatrB and metabolizing enzymes. Frontiers in Plant Science, 14, 1177060.

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Brassinin and Immunoblotting Assay

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Brassinin (catalog no. SML 1635) was purchased from Sigma-Aldrich (St. Louis, MO, USA) and dissolved in dimethyl sulfoxide (DMSO). The antibodies used in the immunoblotting assays are listed in Table 1.

Hong T., Ham J., Song J., Song G, & Lim W. (2021). Brassinin Inhibits Proliferation in Human Liver Cancer Cells via Mitochondrial Dysfunction. Cells, 10(2), 332.

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