All procedures to prepare the nutrient sources are described in Yamazaki and Fujiwara 2022 . Stock solutions of 2 metre NH4Cl and NaH2PO4 · 2H2O (Fujifilm Wako) were used as nutrient sources at a final concentration of 200 mM. To the NaH2PO4 · 2H2O stock solution, 1.07 metre NaOH was added to adjust the pH to 7.4. We supplemented the nutrient sources with N-1-naphthylphthalamic acid (NPA; Naptalam Standard, Fujifilm Wako) at a final concentration of 200 µM, 4-phenoxyphenylboronic acid (PPBo, Tokyo Chemical Industry Co., Ltd, Tokyo, Japan) at 50 µM, (aminooxy)acetic acid hemihydrochloride (AOA, Fujifilm Wako) at 2 mM, paclobutrazol (PBZ, Tokyo Chemical Industry) at 2 mM or 1-aminobenzotriazole (ABT, Tokyo Chemical Industry) at 2 mM, all prepared first as stock solutions in DMSO. NPA was added as an inhibitor of auxin transport via PINs or ABCBs. PPBo, AOA, PBZ and ABT were added as inhibitors of the biosynthesis of auxins, ethylene, gibberellins and salicylic acid via inhibition of YUCCAs, 1-aminocylopropane-1-carboxylic acid synthase, P450 mono-oxygenase and benzoic acid 2-hydroxylase, respectively. In the mock condition, DMSO was added at 1% (v/v) instead of the stock solutions. All stock solutions were stored at −20°C. Nutrient sources were stored at 4°C and used within 1 month of preparation without inhibitors or 1 week with inhibitors (except that PBZ was used within 1 day).
Nah2po4
Manufactured by Fujifilm
Sourced in Japan
NaH2PO4 is a chemical compound commonly used in laboratory settings. It is a white crystalline powder that serves as a source of phosphate ions and sodium ions. This compound is widely utilized in various analytical and experimental procedures due to its chemical properties and versatility.
Automatically generated - may contain errors
Lab products found in correlation
Na2hpo4, by Fujifilm (7 mentions)
Sodium chloride (nacl), by Fujifilm (3 mentions)
Mgso4, by Fujifilm (2 mentions)
Cacl2, by Fujifilm (2 mentions)
Percoll, by GE Healthcare (1 mentions)
K2hpo4, by Fujifilm (1 mentions)
Sodium ascorbate, by Fujifilm (1 mentions)
Beef extract, by MP Biomedicals (1 mentions)
Tween 80, by Fujifilm (1 mentions)
Kh2po4, by Fujifilm (1 mentions)
Nano3, by Fujifilm (1 mentions)
Dmrxa rd, by Leica (1 mentions)
4 6 diamidino 2 phenylindole dapi, by Fujifilm (1 mentions)
Uv 1200, by Shimadzu (1 mentions)
Cholesterol, by Merck Group (1 mentions)
Phosphate buffered saline (pbs), by Nacalai Tesque (1 mentions)
Bodipy pc, by Thermo Fisher Scientific (1 mentions)
Hanks balanced salt solution (hbss), by Nissui Pharmaceutical (1 mentions)
Texas red dppe, by Thermo Fisher Scientific (1 mentions)
Pipes, by Dojindo Laboratories (1 mentions)
12 protocols using nah2po4
1
Nutrient Sources with Inhibitor Supplements
2
Anaerobic Bacteria Cultivation Protocol
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Reagents included De Man Rogosa and Sharpe (MRS) media (Becton Dickinson Difco, U.S.A.), hipolypeptone (Nihon Seiyaku, Japan), beef extract (MP Biomedicals, LLC, France), yeast extract (Becton Dickinson Difco), TBO (Waldeck, Munster), and Percoll (GE Healthcare Life Sciences, Japan). Glucose, Tween 80, K2HPO4, sodium ascorbate, L-cysteine-HCl, NaNO3, MgSO4, KH2PO4, NaH2PO4, NaCl, and 4′,6-diamidino-2-phenylindole (DAPI) were procured from Fujifilm Wako Pure Chemical Corporation, Japan. Data were collected using a spectrophotometer (UV-1200, Shimadzu, Japan) and a fluorescence microscope DMRXA/RD (Leica Microsystems, Germany). Cultures were prepared using fixed-type (model of rotor: BN 4–6) (H-201FR, Kokusan, Japan) and swing-type (model of rotor: RF 110) centrifuges (H-500FR, Kokusan). Deionised and doubly distilled water was used in all experiments.
3
Lipid Characterization and Fluorescent Labeling
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Porcine brain SM; synthetic DOPC, DSPC, DMPC, and DOPE; and commercially available fluorescent SMs were purchased from Avanti Polar Lipids, Inc. Cholesterol and NEG were obtained from Sigma-Aldrich. Stearoyl-SM was purified from the Porcine brain SM by HPLC, and the purity was monitored by thin-layer chromatography. These samples were dissolved at 1 mg/ml in CHCl3/MeOH (4:1 vol/vol) and stored at −20°C until use. ATTO488 and ATTO594 were purchased from ATTO-TEC, Texas-red-DPPE and Bodipy-PC were from Invitrogen, and CTMR was from Thermo Fisher Scientific. These dye compounds were stored in the dark at −20°C until use. PBS was obtained from Nacalai Tesque, NaH2PO4 and Na2HPO4 were from Wako Pure Chemical Industries, HBSS was from Nissui, Pipes was from Dojindo, and Triton X-100 was from ICN Biomedicals. Other chemicals were purchased from Nacalai Tesque.
4
Circular Dichroism Spectroscopy Protocol
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Circular dichroism (CD) measurements were carried out using a Jasco J-820 spectropolarimeter with a 0.1-cm-path-length cuvette at room temperature under constant nitrogen gas flow. Samples were prepared at 0.3 mg/mL in 10 mM phosphate buffer (1.34 g/L Na2HPO4 [Wako Chemicals, Osaka, Japan], adjusted to pH 7.5 with NaH2PO4 [Wako Chemicals]). CD spectra were recorded from 190 to 260 nm at an interval of 0.2 nm and a scan speed of 200 nm/min. Each spectrum was the average of four successive scans.
5
Microplate Reader with Reagent Injectors
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The Nephelostar Plus microplate reader with 2 built-in Reagent injectors and MARS software was obtained from BMG Labtech (Offenburg, Germany) and 96-well plates and 96-well plastic covers were from Corning (Kennebunk, ME, USA). All chemicals (NaCl, Tris, Hepes, CaCl2, NaH2PO4, Na2HPO4, NaOH, ZnCl2) were pro-analysis -grade quality and purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan).
6
Optimization of BiSCaO-6 Suspension pH
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Addition to 0.2 g BiSCaO-6 to 100 mL of pure water, followed by rotary mixing, generated 0.2 wt% BiSCaO-6 suspensions. First, various amounts of phosphoric acid (H3PO4), hydrochloric acid (HCl) or sulfuric acid (H2SO4) were added to 0.2 wt% BiSCaO-6 suspensions to adjust the pH to around 6, 7, 8, 9, 10.5, 12, and 12.5. After the measurement of pH with a pH meter (F-70, HORIBA Ltd., Kyoto, Japan), the form and the proportion of dispersion or flocculation to total amount were observed. Next, 0.04 wt%, 0.12 wt%, 0.2 wt%, and 0.28 wt% of Na3PO4, Na2HPO4, or NaH2PO4 (FUJI FILM Wako Pure Chemical Corp.) were added to 0.2 wt% BisCaO-6 water suspension, rotary mixed, a7nd then evaluated for pH, form, and the proportion of layer separation with flocculation to the total amount.
7
Recording Neuronal Activity in Mice Brain Slices
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C57BL/6NCrSlc mice (Six weeks old) were obtained from Japan SLC. Inc. Slices of mice brain were prepared as 300 µm using a slicer (NeoLinearSlicer MT, Dosaka EM). The study was approved by Tohoku Institute of Technology Animal Care and User Committee (approval number: 2020‐01). Brain slices were plated on CMOS‐MEA supplemented with 0.05% polyethyleneimine (Sigma), incubated for 1 h, then washed with DW and dried overnight. During the experiments, the CMOS‐MEA were perfused with artificial cerebrospinal fluid (in [× 10‐3m ]: NaCl (KCH5696, FUJIFILM) 124, KCl (160‐03555, Wako) 3, NaH2PO4 (169‐04245, Wako) 1.25, MgSO4 (131‐00405, Wako) 0.1, Glucose (044‐00695, Wako) 10, NaHCO3(191‐01305, Wako) 26, CaCl2(039‐00475, Wako) 3.0 with carbogen (95% O2, 5% CO2) at a flow rate of 1.5 mL min‐1 using a perfusion device [(Gilson, Inc.) Mini Pulse Pump III MP‐4]. The local field potential (LFP) of each brain region was measured at 1 kHz sampling rate with 236880 electrodes simultaneously.
8
Enzymatic Hydrolysis of Cellulosic Biomass
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Chemicals. Deionized water, purified using a Milli-Q® Advantage A10® system (Millipore™, Eschborn, Germany), was used. Phosphate buffer was prepared from Na2HPO4 and NaH2PO4 purchased from Wako Pure Chemical Industries. The enzyme cocktail was prepared by combining equal amounts of commercial enzymes OPTIMASH XL containing cellulase and xylanase (10,300 U/g) and OPTIMASH BG containing xylanase and β-glucosidase (6200 U/g) from DuPont™ Genencor® Science. Jet milled cedar (18 μmΦ) was used for lab-scale and large milling operations.
9
VHH-display Phage Binding Assay
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Each round of VHH-display phage was prepared according to the aforementioned procedure. One hundred microliters of FGFR1β (IIIc)/Fc (10 μg/ml) was added to Immuno Clear Standard Modules, and the plate was left at 4 °C overnight. After washing with PBS three times, the PBS containing 3% skim milk was added to wells and incubated at RT for 1 h. Each well was washed with PBS-T four times. The 50 μl of each round of VHH-display phage in PBS, which contained 10% BSA, was added to each well and incubated at RT for 1 h. Each well was washed with PBS-T four times. About 50 μl of Anti-M13-mAb-horseradish peroxidase in PBS (1/3000 dilution; Sino Biological) was added and incubated at RT for 1 h. After washing with PBS five times, OPD detection reagent (1 tablet in 0.1 M NaH2PO4 plus 0.01% H2O2; Fujifilm Wako Pure Chemical Corporation) was added to each well and allowed to develop in the dark. After enough color development, 1 M H2SO4 was added, and absorbance was measured at 490 nm (reference; 630 nm) using Infinite M Plex (Tecan Group Ltd).
10
Sodium Phosphate Compounds Characterization
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NaH 2 PO 4 (Japanese pharmaceutical excipients), NaH 2 PO 4 ▪2H 2 O (Guaranteed reagent), Na 2 HPO 4 (for pH standard solution and Japanese pharmaceutical excipients) and Na 2 HPO 4 ▪12H 2 O (Guaranteed reagent) were purchased from Fujifilm Wako Pure Chemical Corporation (Osaka, Japan). NaH 2 PO 4 ▪H 2 O (ACS reagent) and Na 2 HPO 4 ▪H 2 O (ACS reagent) were obtained from Correspondence: Toshiyuki Ohtake (E-mail: toshiyuki.otake@shiseido.com) Sigma-Aldrich Co. LLC (St. Louis, MO, USA). A 20 mM sodium phosphate buffer (NaPB, pH7.4) was prepared using ultrapure water (Fujifilm Wako) as described in OECD TG 495 (OECD, 2019) .
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