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15nh415no3

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15NH415NO3 is a chemical compound used in laboratory settings. It is comprised of nitrogen, hydrogen, and oxygen atoms in a specific molecular structure. This product serves as a tool for researchers and scientists, providing a standardized chemical substance for various experimental and analytical applications.

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Lab products found in correlation

K15no3, by Merck Group (2 mentions) Isoprime, by Elementar (1 mentions)

6 protocols using 15nh415no3

1

Stable Isotope Labeling of Oak Microcuttings

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Twenty-four hours before harvest, plants were transferred into a plexiglas chamber and labelled using a mobile 13CO2 labelling system controlling CO2 concentration and air humidity (set to 400 µl l−1 and 70%, respectively) in the chamber (see [26 (link),31 ]). CO2 in ambient air was removed and replaced by CO2 containing 8.3 ± 0.2 atom% 13CO2 (mean ± s.d.) (Eurisotop, Saarbrücken, Germany). Thereby, Q. robur microcuttings were exposed to the 13CO2-enriched atmosphere over a complete light period of 16 h and the CO2 concentration adjusted to 400 ± 2 ml l−1 as described in Herrmann et al. [30 (link)]. 15N labelling was performed 72 h before harvest using 98 atom% 15NH415NO3 (Sigma, Darmstadt, Germany); 0.1 mg 15NH415NO3 dissolved in 5 ml sterile distilled water was injected into the rhizosphere under sterile conditions.
Graf M., Bönn M., Feldhahn L., Kurth F., Grams T.E., Herrmann S., Tarkka M., Buscot F, & Scheu S. (2019). Collembola interact with mycorrhizal fungi in modifying oak morphology, C and N incorporation and transcriptomics. Royal Society Open Science, 6(3), 181869.
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2

Isotopic Labeling of Arabidopsis Proteins

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Metabolic labeling of proteins in Arabidopsis was performed according to the protocol reported previously (Skirycz et al., 2011 ). The 1/2 MS media were prepared to contain 14N and 15N isotopes. In the 15N-incorporated medium, the heavy labeling reagents of 15NH415NO3 and K15NO3 (Sigma) were used to replace those containing 14N-isotope. Arabidopsis seedlings were grown at 22°C under long-day conditions (16 : 8 h, light : dark) on 15N- or 14N-isotope-containing medium, respectively. After 2 wk, the seedlings grown on the 14N medium were transferred to a growth chamber at 4°C for 6, 12 and 18 d. As such, 15N-labeled plants at 22°C were used for control experiments (i.e. forward labeling), or the time-course cold-treated plants at 4°C were also metabolically labeled by 15N-isotopes in parallel experiments (i.e. reciprocal labeling) (Kline et al., 2010 ; Qing et al., 2016 (link)). The labeled and unlabeled seedlings were harvested separately, and a mixture with the same amount of 14N- and 15N-isotope-labeled tissues (w/w, 1:1) at the two different temperatures was utilized for relative quantification of proteins.
Ma J., Wang D., She J., Li J., Zhu J.K, & She Y.M. (2016). Endoplasmic reticulum-associated N-glycan degradation of cold-upregulated glycoproteins in response to chilling stress in Arabidopsis. The New phytologist, 212(1), 282-296.
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3

Quantifying Plant Nitrogen Uptake with 15N Enrichment

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To measure N uptake by plants, 15N-enriched ammonium nitrate was applied once in each pot at 1 month post-inoculation (15NH415NO3, > 98% 15N; Sigma Aldrich, St. Louis, United States) to reach δ 15N = + 4450, corresponding to an enrichment of 2% (calculated value using isotopic abundance of the unlabeled and 15N-enriched ammonium nitrate and their molar ratio in the liquid fertilizer). Dry shoot and root materials were ground separately in tungsten tubes using tungsten carbide balls in a Retch MM301 vortexer (Retch GmbH & Co., Haan, Germany). Total N, 15N and C aliquots of 2 mg were weighed for elemental analyses, and their concentrations were determined using an ANCA elemental analyzer/mass spectrometer (Wellience, Dijon, France). To measure phosphorus (P) content of plants, dry leaf material (20 mg) was ashed at 550°C for 8 h. The residue was dissolved in 2 ml H2O and 100 μl HCl (32%). One ml of the solution was then transferred to a 2-ml Eppendorf tube and centrifuged at 12,000 rpm for 10 min in a benchtop centrifuge. One hundred μl of the supernatant were transferred into a well of a 96-wells microtiter plate. Phosphorus concentration was then measured by the malachite green method (Ohno and Zibilske, 1991 (link)).
García-Soto I., Boussageon R., Cruz-Farfán Y.M., Castro-Chilpa J.D., Hernández-Cerezo L.X., Bustos-Zagal V., Leija-Salas A., Hernández G., Torres M., Formey D., Courty P.E., Wipf D., Serrano M, & Tromas A. (2021). The Lotus japonicus ROP3 Is Involved in the Establishment of the Nitrogen-Fixing Symbiosis but Not of the Arbuscular Mycorrhizal Symbiosis. Frontiers in Plant Science, 12, 696450.
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4

Arabidopsis Protein Turnover Kinetics

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Seven-day-old Arabidopsis cell culture was transferred from non-labelled media (14N) to media without nitrogen, and the cells washed three times to eliminate 14N in the media. Washed cells were transferred to heavy media (15N) containing two nitrogen sources for optimal growth [16 (link)], namely 1.65 g/l 15NH415NO3 (Sigma–Aldrich 299278) and 1.9 g/l K15NO3 (Sigma–Aldrich 335134). To study protein turnover rate, the labelled Arabidopsis cell culture was collected by vacuum filtration after the first day, third day and fifth day following transfer into new media, then stored at −80°C until use.
Salih K.J., Duncan O., Li L., Trösch J, & Millar A.H. (2020). The composition and turnover of the Arabidopsis thaliana 80S cytosolic ribosome. Biochemical Journal, 477(16), 3019-3032.
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5

Tracking C and N Allocation in Plant-Microbe Interaction

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The allocation patterns of C and N were assessed by stable isotope labelling. Three days before harvesting, 5 ml of 0.02 g l−1, 15NH415NO3 (98 atm% 15N, Sigma, Germany) were added to each root compartment under sterile conditions. For 13CO2 labelling, microcosms were transferred into a Plexiglas chamber 36 h before harvest. During the night before labelling, the CO2 in the atmosphere was scrubbed with soda lime and replaced with CO2 with 10 atm% 13CO2 (Eurisotop, Saarbrücken, Germany). During the following daytime period (16 h), the CO2 concentration was adjusted to 400 ± 2 μl l−1 (mean ± SD) with a 13C atm% of 7.9 ± 0.3 (mean ± SD). Carbon isotopic composition of atmospheric samples was assessed using an isotope ratio mass spectrometer (Isoprime; Elementar, Hanau, Germany). For further details see23 . In the used culture system, we expect that the 13C and 15N excess values result from the assimilation by the plant. The 13CO2 is assimilated by the plant, and none of the interacting organisms can fix atmospheric 13CO2. The 15N is taken up by the roots, and not by the ectomycorrhizal fungus. This is, because in the used culture system only few ectomycorrhizas formed (< 1% of fine root biomass). Thus, we interpret changes in N and C allocation as an indication of plant response to the interacting organism.
Tarkka M.T., Grams T.E., Angay O., Kurth F., Maboreke H.R., Mailänder S., Bönn M., Feldhahn L., Fleischmann F., Ruess L., Schädler M., Scheu S., Schrey S.D., Buscot F, & Herrmann S. (2021). Ectomycorrhizal fungus supports endogenous rhythmic growth and corresponding resource allocation in oak during various below- and aboveground biotic interactions. Scientific Reports, 11, 23680.
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6

Arabidopsis thaliana Growth Optimization

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Arabidopsis thaliana Col-0 seeds were sown on 0.8% agar / ddH 2 O (w/v) contained in 2 ml Eppendorf tube lids punctured with ~3mm diameter holes. Lids were then placed into 24 tube racks and vernalised before transferring to 12 / 12 hour day (220 𝜇mol/m 2 /second white light) night cycle at a constant 22°C. 0.5 strength Hoagland's media was used throughout with solutions being changed weekly. Reagents including heavy isotopes were purchased from Sigma 5% 2 H 2 O supplemented solutions were made up v/v with 99.9% deuterium oxide while 15 N solutions contained K 15 NO 3 98% and 15 NH 4 15 NO 3 98% obtained from Sigma Aldrich.
Duncan O., & Millar A.H. (2021). BREAKTHROUGH REPORT - Large scale measurement of protein synthesis rates over the diurnal cycle reveal pathway specific regulation of translation.
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