Nahco3

Manufactured by Nacalai Tesque

Sourced in Japan

Sodium bicarbonate (NaHCO3) is a white crystalline powder commonly used as a laboratory chemical. Its core function is to act as a buffer, maintaining a stable pH in various chemical reactions and processes.

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Lab products found in correlation

18 protocols using nahco3

Surface Modification and Characterization of Titanium Alloys

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A commercially pure, medical-grade Ti plate (ISO5832-2) (Ti > 99.5%) was provided by Nilaco Co., Tokyo, Japan. Ti-6Al-4V alloy plate (Ti = balance, Al = 6.18, V = 4.27 mass%) and Ti-15Zr-4Nb-4Ta alloy plate (Ti = balance, Zr = 14.51, Nb = 3.83, Ta = 3.94, Pd = 0.16, and O = 0.25 mass%) were supplied by Kobelco Research Institute, Inc., Hyogo, Japan.
The chemical reagents (NaOH, CaCl₂, ICl₃, ICl, NaI and povidone iodine (PVP-I)) used for surface treatment were reagent-grade and purchased from Kanto Chemical Co., Inc., Tokyo, Japan. Reagent-grade NaCl, NaHCO₃, KCl, K₂HPO₄·3H₂O, MgCl₂·6H₂O, CaCl₂, Na₂SO₄, tris-hydroxymethylaminomethane (CH₂OH)₃CNH₂, and 1 M HCl were purchased from Nacalai Tesque, Inc., Kyoto, Japan, and used for the preparation of simulated body fluid (SBF). Minimum essential medium (MEM) was obtained from Gibco, Thermo Fisher Scientific, Waltham, MA, USA, and used for the cell culture test. Nutrient broth (Eiken Chemical Co., Ltd. Tochigi, Japan) and RPMI 1640 (Fujifilm Wako Pure Chemical Corp., Osaka, Japan) were used for the antibacterial activity test.

Yamaguchi S., Le P.T., Shintani S.A., Takadama H., Ito M., Ferraris S, & Spriano S. (2021). Iodine-Loaded Calcium Titanate for Bone Repair with Sustainable Antibacterial Activity Prepared by Solution and Heat Treatment. Nanomaterials, 11(9), 2199.

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Heterohybridoma Generation and Culture

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Three gerbil–mouse heterohybridomas—B11D2(C2)(which secrete gerbil IgG1 specific to
keyhole limpet hemocyanin; KLH) [21 (link), 22 (link)], D9(E4), and D9(E6)C2B3, which were generated by
fusing gerbil splenocytes with mouse myeloma cells (P3-X63-Ag8.653, provided by the RIKEN
BioResource Center)—and mouse myeloma cells (P3-X63-Ag8.653) were cultured in RPMI-1640
medium (Wako, Osaka, Japan) supplemented with 10% heat-inactivated fetal bovine serum (JRH
Biosciences, Tokyo, Japan), 100 U/ml penicillin (Meiji, Tokyo, Japan), 100
µg/ml streptomycin (Meiji), MEM nonessential amino acids (Invitrogen
Gibco, Tokyo, Japan), 5 × 10⁻² M 2-mercaptoethanol (Wako, Tokyo, Japan), and 2
µg/ml NaHCO₃ (Nacalai Tesque, Tokyo, Japan). Cells were
maintained in a humidified incubator at 37°C with 5% CO₂. Media were changed
three times a week.

UKAJI T., HASHIMOTO M, & KAI O. (2015). Conditioned medium from gerbil—mouse T cell heterohybridomas improved antibody secretion. Experimental Animals, 64(2), 199-205.

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In Vitro Evaluation of Bioactive Polymer Screw HA Formation

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The hydroxyapatite (HA)-forming ability of bioactive PSs was examined by incubation in simulated body fluid (SBF) as previously reported[26 (link)]. Briefly, samples were soaked in 24 ml of SBF at 36.5°C with the following ion concentrations approximating those of human blood plasma: Na⁺, 142.0; K⁺, 5.0; Ca²⁺, 2.5; Mg²⁺, 1.5; Cl^–, 147.8; HCO₃^–, 4.2; HPO₄^2-, 1.0; and SO₄^2-, 0.5 mM. The SBF was prepared by dissolving reagent grade NaCl, NaHCO_3, KCl, K₂HPO₄·3H₂O, MgCl·6H₂O, CaCl₂, and Na₂SO₄ (Nacalai Tesque Inc., Kyoto, Japan) in ultrapure water, buffered at pH = 7.4 with tris (hydroxymethyl) aminomethane (CH₂OH)₃CNH₂ and 1 M HCl (Nacalai Tesque Inc., Kyoto, Japan) at 36.5°C[27 (link)]. After soaking in SBF for three days, the samples were gently rinsed with ultrapure water and dried. HA formation on the surface of screws was evaluated by FE-SEM and energy dispersive X-ray analysis (EDX; EMAX-7000, Horiba Ltd., Kyoto, Japan). HA-forming ability was also evaluated on bioactive PSs that had been removed after insertion into the pedicle of cryopreserved canine lumbar spines.

Akeda K., Yamaguchi S., Matsushita T., Kokubo T., Murata K., Takegami N., Matsumine A, & Sudo A. (2018). Bioactive pedicle screws prepared by chemical and heat treatments improved biocompatibility and bone-bonding ability in canine lumbar spines. PLoS ONE, 13(5), e0196766.

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Apatite Formation on Porous Titanium

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Cylindrical specimens of the porous Ti metals subjected to the various treatments given in Table 1 were broken and soaked in 30 mL of a simulated body fluid (SBF) having ion concentrations close to those of human blood plasma (Na⁺ = 142.0, K⁺ = 5.0, Mg²⁺ = 1.5, Ca²⁺ = 2.5, Cl^– = 147.8, HCO₃^– = 4.2, HPO₄^2– = 1.0, and SO₄^2– = 0.5 mM) [19] (link), [20] (link) at 36.5°C. The SBF was prepared by dissolving reagent-grade NaCl, NaHCO₃, KCl, K₂HPO₄·3 H₂O, MgCl₂.6 H₂O, CaCl₂, and Na₂SO₄ (Nacalai Tesque Inc., Japan). After 1 day, the specimens were removed from the SBF solution, gently washed with ultrapure water and dried in an oven at 40°C. Formation of apatite on their surfaces was examined by SEM and TF-XRD analysis.

Kawai T., Takemoto M., Fujibayashi S., Akiyama H., Tanaka M., Yamaguchi S., Pattanayak D.K., Doi K., Matsushita T., Nakamura T., Kokubo T, & Matsuda S. (2014). Osteoinduction on Acid and Heat Treated Porous Ti Metal Samples in Canine Muscle. PLoS ONE, 9(2), e88366.

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Large-scale AAV vector production

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AAV vectors were prepared in a large scale and harvested from culture supernatant (conditioned media), as previously described [3 (link)]. In brief, a 293EB cell line expressing adenoviral E1a, adenoviral E1b, and Bcl-x_L [15 (link)] was expanded in two 500 mL flasks (HYPERFlask, Corning, Corning, NY, USA) or a 1 L bioreactor (iCELLis Nano Bioreactor, Pall, Port Washington, NY, USA) for 5 days or 4 days, respectively, in Dulbecco’s Modified Eagle Medium (DMEM high glucose, FUJIFILM Wako, Chuo-ku, Osaka, Japan) with 10% fetal bovine serum (Thermo Fisher, Waltham, MA, USA). Transfection was then performed with polyethylenimine max, (Polysciences, Warrington, PA, USA) using pAAV-ZsGreen1 (TaKaRa Bio, Kusatsu, Shiga, Japan), pRC9 (serotype 9), and helper plasmids in DMEM including 2 mM L-Alanyl-L-glutamine Solution(100x) (Nacalai Tesque, Nakagyo-ku, Kyoto, Japan), 0.12% NaHCO₃ (Nacalai Tesque), and 0.13% D-glucose (Nacalai Tesque) without serum. Five days post-transfection, culture supernatants were harvested and treated with 18.5 U/mL endonuclease (KANEKA CORPORATION, Minato-ku, Tokyo, Japan) with 5 mM MgCl₂ (Nacalai Tesque) for 30 min at 37 °C. All cells were checked for mycoplasma contaminations resulting were reported negative.

Wada M., Uchida N., Posadas-Herrera G., Hayashita-Kinoh H., Tsunekawa Y., Hirai Y, & Okada T. (2023). Large-scale purification of functional AAV particles packaging the full genome using short-term ultracentrifugation with a zonal rotor. Gene Therapy, 30(7-8), 641-648.

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Luminol-Based Chemiluminescence Immunoassay

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Luminol, dithiothreitol, bovine serum albumin (BSA), NaCl, NaHCO₃, Na₂CO₃, H₂SO₄, HCl, CH₃COONa, Na₂HPO₄, and CH₃COOH were acquired from Nacalai Tesque (Kyoto, Japan). KCL, tween 20, hydrogen peroxide, p-iodonitrotetrazolium (INT), sodium borohydride, and KH₂PO₄ were sourced from Wako Pure Chemical Industries (Osaka, Japan). Anti-rabbit IgG-biotin conjugate and avidin were obtained from the following USA companies: ImmunoReagent Inc. (Burlington, NC, USA) and Calzyme Laboratories Inc. (San Luis, CA, USA), respectively. Biotin-hydrazide and NaIO₄ were purchased from Sigma-Aldrich (St. Louis, MO, USA), while sodium hydroxide was from Merck (Darmstadt, Germany). Dextran 40 was obtained from TCI (Tokyo, Japan). Doxorubicin HCl was purchased from Funakoshi co (Tokyo, Japan). Double distilled water was used throughout the whole experiment, and it was produced from Yamato Autostill WG203 (Tokyo, Japan).
By combining sodium chloride, potassium dihydrogen phosphate, disodium phosphate, and potassium chloride, phosphate buffer saline (PBS, 100 mM pH 7.4) was prepared. A 50.0 mM carbonate-bicarbonate buffer with pH 9.6 was made from Na₂CO₃ and NaHCO₃.

Kaladari F., Kishikawa N., Shimada A., El-Maghrabey M, & Kuroda N. (2023). Anthracycline-Functionalized Dextran as a New Signal Multiplication Tagging Approach for Immunoassay. Biosensors, 13(3), 340.

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Cellulose Hydrolysis and Characterization

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5‐HMF (>95.0 %, GC) and dimethyl sulfone (DMSO₂, >99.0 %, GC) were purchased from Tokyo Chemical Industry, Tokyo, Japan. Before experiments, water was removed from 5‐HMF by freeze‐drying. Whatman No. 42 filter paper was used as a cellulose sample, which was cut into small sheets (7×7 mm). Glycerol (>99.5 %, GC), Na₂CO₃ (>99.5 %, titration) and NaHCO₃ (99.5 %, titration) were purchased from Nacalai Tesque, Inc., Kyoto, Japan, and used without purification. Aqueous NaOH solution (47–53 %, T) and aqueous HCl solution (35–37 %, T) were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan) and used without purification.

Nomura T., Minami E, & Kawamoto H. (2021). Hydroxymethylfurfural as an Intermediate of Cellulose Carbonization. ChemistryOpen, 10(6), 610-617.

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Simulated Body Fluid Soaking Protocol

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The samples subjected to the MA-heat or NaOH-HCl-heat treatments were soaked in 24 mL of a simulated body fluid (SBF) with various ion concentrations (Na⁺ 142.0, K⁺ 5.0, Ca²⁺ 2.5, Mg²⁺ 1.5, Cl⁻ 147.8, HCO₃⁻ 4.2, HPO₄²⁻ 1.0, and SO₄²⁻ 0.5 mM) closed to human blood plasma at 36.5 °C. The SBF was prepared by dissolving reagent grade NaCl, NaHCO₃, KCl, K₂HPO₄•3H₂O, MgCl₂•6H₂O, CaCl₂, and Na₂SO₄ (Nacalai Tesque Inc., Kyoto, Japan) in ultrapure water, and buffered at pH = 7.4 with tris(hydroxymethyl)aminomethane (CH₂OH)₃CNH₂ and 1 M HCl (Nacalai Tesque Inc., Kyoto, Japan) at 36.5 °C [8 (link)]. After soaking in the SBF for 3 and 7 days, the samples were gently rinsed with ultrapure water, and dried.

Yamaguchi S., Hashimoto H., Nakai R, & Takadama H. (2017). Impact of Surface Potential on Apatite Formation in Ti Alloys Subjected to Acid and Heat Treatments. Materials, 10(10), 1127.

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Quantifying Intracellular Reactive Oxygen Species

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Intracellular ROS were measured using the DCFDA Cellular ROS Detection Assay Kit (Abcam) with the instructions provided by the manufacturer. Briefly, on day 9 of treatment of 3HBA on 3T3-L1 adipocytes (Fig. 4), or on day 8 of transfection with small interfering RNA (Fig. 7), culture medium was replaced with Krebs–Ringer Bicarbonate buffer (KRBB), composed of 25 mM NaHCO₃ (Nacalai), 119 mM NaCl (Nacalai), 4.74 mM KCl (Nacalai), 1.19 mM MgCl₂ (Nacalai), 1.19 mM KH₂PO₄ (Nacalai), 2.54 mM CaCl₂ (Nacalai), 10 mM 4-(2-hydroxyethyl)-1-iperazineethanesulfonic acid (Nacalai), and 0.05 mM Bovine Serum Albumin solution (Sigma), supplemented with 10 μM DCFDA. After staining with DCFDA for 1 h at 37 ℃ under 5% CO₂, 3T3-L1 adipocytes were wash 3 times with PBS and detected by fluorescence spectroscopy with excitation / emission at 485 nm / 535 nm.

Nishitani S., Fukuhara A., Tomita I., Kume S., Shin J., Okuno Y., Otsuki M., Maegawa H, & Shimomura I. (2022). Ketone body 3-hydroxybutyrate enhances adipocyte function. Scientific Reports, 12, 10080.

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Bovine Oocyte In Vitro Fertilization

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After in vitro maturation, the COCs were transferred into a 15 ml tube containing 2 ml of HEPES-TLP-PVA supplemented with 0.1% (w/v) hyaluronidase (Sigma) and vortexed for
90 sec to remove cumulus cells from oocytes. Denuded oocytes were transferred to droplets (80 µl) of fertilization medium in groups of 15 to 20 under paraffin oil in a 35 mm polystyrene
dish. The fertilization medium was composed of 114.0 mM NaCl (Nacalai), 3.2 mM KCl (Nacalai), 6.76 mM CaCl₂·2H₂O (Nacalai), 0.5 mM MgCl₂·6H₂O
(Nacalai), 0.1 mM sodium pyruvate, 10.0 mM sodium lactate (Sigma), 0.35 mM NaH₂PO₄·2H₂O (Nacalai), 5.0 mM D-glucose, 25.0 mM NaHCO₃ (Nacalai),
0.3% (w/v) bovine serum albumin (Fraction V; Sigma), 100 µg/ml amikacin sulfate, and 2.0 mM caffeine (Sigma). The frozen spermatozoa were thawed immediately before insemination, as described
above. HEPES-TLP-PVA containing frozen-thawed spermatozoa was centrifuged at 760 ×g for 10 min at 38°C, and the supernatant was removed. The precipitated spermatozoa were
gently suspended in the fertilization medium at a concentration of 3.5 × 10⁷ spermatozoa/ml, and 20 µl of this sperm suspension was introduced into the 80 µl droplet that
contained denuded oocytes at a final concentration of 7.0 × 10⁶ spermatozoa/ml. The oocytes and spermatozoa were then cultured for 12 h at 38.5°C in an atmosphere with 5%
CO₂ in air.

JIN H., CHOI W., MATSUMURA K., HYON S.H., GEN Y., HAYASHI M., KAWABATA T., IJIRI M, & MIYOSHI K. (2022). Cryopreservation of pig spermatozoa using carboxylated poly-L-lysine as cryoprotectant. The Journal of Reproduction and Development, 68(5), 312-317.

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