Dbmib

Manufactured by Merck Group

Sourced in United States

DBMIB is a laboratory instrument used for the measurement and analysis of biological samples. It functions as a spectrophotometer, capable of detecting and quantifying the presence of specific compounds within a sample.

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Lab products found in correlation

N propyl gallate, by Merck Group (1 mentions) Nigericin, by Nacalai Tesque (1 mentions) Antimycin a, by Merck Group (1 mentions) Dm2500, by Leica (1 mentions) Atrazine, by Merck Group (1 mentions)

6 protocols using dbmib

Probing PSII Electron Transfer with PTOX Inhibitors

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To determine the contribution of the PTOX to the entire photosystem II (PSII) electron transfer, the leaves of either untreated (control) or salt-treated SV and SA were vacuum infiltrated with either water or with 5 mM n-propyl gallate (n-PG, 3,4,5-trihydroxy-benzoic acid-n-propyl ester; Sigma) or 50 μM DBMIB (2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone, Sigma). The stock solutions of n-PG were freshly prepared in ethanol and DBMIB in methanol.
To determine whether PTOX may play a role in electron transfer from PSII to O₂, measurements of ETR_II were performed on leaves obtained from control and salt-treated SA and SV which were vacuum infiltrated with either water or a solution of the PTOX inhibitor n-propyl gallate (n-PG; 3,4,5-trihydroxy-benzoic acid-n-propyl ester; Joët et al., 2002 (link); Josse et al., 2003 (link); Kuntz, 2004 (link); Rosso et al., 2006 (link); Houille-Vernes et al., 2011 (link); Sun and Wen, 2011 (link); Trouillard et al., 2012 (link); Shirao et al., 2013 (link); Nawrocki et al., 2015 (link)).

Essemine J., Lyu M.J., Qu M., Perveen S., Khan N., Song Q., Chen G, & Zhu X.G. (2020). Contrasting Responses of Plastid Terminal Oxidase Activity Under Salt Stress in Two C4 Species With Different Salt Tolerance. Frontiers in Plant Science, 11, 1009.

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Photosynthesis Fluorescence Kinetics in Cyanobacteria

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Reaction center closure was followed using a PSI ﬂuorometer (PSI Instruments, Brno, Czech Republic) in the 1-ms to 1-s time range, in dark-adapted (15 min) and quenched Synechocystis cells. The concentration of cells was set to 2.5 μg Chl/mL. WT and ΔapcC cells were illuminated with 1,200 μmol photons m⁻² s⁻¹ of blue–green light during 3 min to obtain quenched cells. Before measuring, DCMU (10 μM) and DBMIB (20 μM; Sigma) were added. Blue measuring light (λ = 460 nm) and orange actinic light (35 μmol photons m⁻² s⁻¹, λ = 630 nm) were used in all cases.

Zhan J., Steglich C., Scholz I., Hess W.R, & Kirilovsky D. (2020). Inverse regulation of light harvesting and photoprotection is mediated by a 3′-end-derived sRNA in cyanobacteria. The Plant Cell, 33(2), 358-380.

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Verification of Electron Transfer Inhibitors

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The inhibitors DCMU (Sigma, United States), DBMIB (Sigma, United States) were prepared as standard in ethanol. We purchased two separate stocks of DNP-INT (caymann chemicals batch no. 0468094-2, 0468093-2, 0524164-2) with the second being sent for analytical analysis to confirm purity via ¹HNMR. In addition we received another DNP-INT stock from the original Trebst preparation (donated by Anja Krieger). All DNP-INT stock solutions were prepared in dimethyl sulfoxide and used separately to repeat experiments as stated in the text. For conducting each experiment, all stock solutions used were freshly prepared and the concentrations of all inhibitors used were based on those experimentally demonstrated in literature to completely block electron transfer process.

Fitzpatrick D., Aro E.M, & Tiwari A. A. (2020). A Commonly Used Photosynthetic Inhibitor Fails to Block Electron Flow to Photosystem I in Intact Systems. Frontiers in Plant Science, 11, 382.

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Exploring Photosynthetic Inhibitors' Effects

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DCMU, DBMIB, ammonium chloride, and Antimycin A were purchased from Sigma–Aldrich and Nigericin was purchased from Nacalai Tesque Inc. To detect responsiveness to light, leaf discs were vacuum-infiltrated in the dark with a solution containing 10 or 100 µM DCMU, 10 or 100 µM DBMIB, 5 or 50 mM ammonium chloride, 10 mM DTT, 1 mM or 10 mM NaF and 50 µM Nigericin for at least 10 min before illumination. As a control, leaves were treated with 1% (v/v) ethanol (used as a solvent for the inhibitors). For isolated chloroplasts, equal volumes of solution containing 50 mM HEPES/KOH, 10 mM MgCl₂, 0.3 M sorbitol, and 10 mM NAD⁺ with 20 µM DCMU, 20 µM DBMIB, 10 mM ammonium chloride, or 10, 50, or 100 µM Antimycin A was added to the chloroplast suspension before illumination. Dark-response experiments were initiated immediately after the addition of an equal volume of solution containing 20 or 100 µM DCMU, 10 mM ammonium chloride, or 100 µM Antimycin A to 1% (v/v) ethanol in which the leaf discs were floating under light. Thereafter, leaf discs were incubated under either light or dark conditions.

Fukuda Y., Ishiyama C., Kawai-Yamada M, & Hashida S.N. (2023). Adjustment of light-responsive NADP dynamics in chloroplasts by stromal pH. Nature Communications, 14, 7148.

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Photosynthesis and Sporulation Dynamics

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To study how photosynthetic electron transport influences sporulation and spore release, the photosynthetic inhibitors 3-(3′,4′-dichlorophenyl)-1,1-dimethylurea (DCMU) (Sigma Aldrich) and dibromothymoquinone (DBMIB) (Sigma Aldrich) were used to treat the 0.75 mm disks cultured in common sterile seawater respectively. The control was the normal group without DCMU or DBMIB. The final concentrations of DCMU and DBMIB were 10 μM. In order to prevent DBMIB to act as a PS II electron acceptor, we used 1 mM ascorbate to keep DBMIB reduced34 . The DCMU treatments were conducted in sets of disks designated as groups 1, 2, 3, and 4, which had been previously cultured for 0, 24, 36, and 48 h, respectively. After treatment with the inhibitor, these groups continued to be cultured until reaching a total of 84 h. The DBMIB treatments followed the same protocol. The status of these groups was regularly monitored and recorded by microscope (Leica DM2500, Germany).

Wang H., Lin A., Gu W., Huan L., Gao S, & Wang G. (2016). The sporulation of the green alga Ulva prolifera is controlled by changes in photosynthetic electron transport chain. Scientific Reports, 6, 24923.

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Cyanobacterial Methane Production Inhibition

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Three photosynthesis inhibitors were used to observe their effect on cyanobacterial CH₄ production. All three inhibitors were dissolved in water rather than ethanol as the latter results in an ionization pattern that masks that of CH₄. Atrazine (catalog number 45330-250MG-R, Sigma-Aldrich) has a solubility of 33 mg liter⁻¹ (153 μM) in water and was used at final concentrations of 5 and 10 μM. DBMIB (catalog number 271993-250MG, Sigma-Aldrich) has a solubility of 132 mg liter⁻¹ (410 μM) and was used at final concentrations of 5, 10, and 40 μM. HQNO (catalog number sc-202654A, Santa Cruz Biotechnology) was described as only very slightly soluble in water without any numerical information. We used culture (0.1 mg ml⁻¹) yet noticed that the powder grains remained nearly intact. Nevertheless, given an observed effect on the culture, we concluded that part of the material dissolved in the culture media. For the experiments, cultures of Anabaena sp. PCC 7120 were concentrated and resuspended in fresh BG11 medium and incubated in a 10-ml analysis chamber connected via circular flow to the MIMS. Inhibitors were introduced via injection of a concentrated solution at 11:00 after 24 hours from the experiment start to allow the culture to acclimate. The light regime used was the same as described above.

Bižić M., Klintzsch T., Ionescu D., Hindiyeh M.Y., Günthel M., Muro-Pastor A.M., Eckert W., Urich T., Keppler F, & Grossart H.P. (2020). Aquatic and terrestrial cyanobacteria produce methane. Science Advances, 6(3), eaax5343.

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