All procedures involving animals were conducted in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals following protocols approved by the University of California, Davis Institutional Animal Care and Use Committee. Timed-pregnant Sprague Dawley rats were purchased from Charles River Laboratory (Hollister, CA). All animals were housed in clear plastic shoebox cages containing corn cob bedding under constant temperature (22 ± 2 °C) and a 12-h light-dark cycle. Food and water were provided ad libitum. Primary cortical cell cultures were prepared from postnatal day 0 rat pups as previously described [26 (link)]. Neocortices from all pups in the litter were pooled, dissociated, and plated at a density of 650 cells/mm2 on substrates precoated with 0.5 mg/mL of poly-l -lysine (Sigma) in B buffer (3.1 mg/mL boric acid and 4.75 mg/mL borax, Sigma) for 4 h at 37 °C and 5% CO2 then washed with sterile deionized water and covered with plating medium. Primary cortical cells were plated in plating medium and allowed to adhere for 4 h before the medium was changed to the co- or tri-culture medium. Half-media changes were performed at DIV 3, 7, and 10 with the respective media types.
Borax
Manufactured by Merck Group
Sourced in United States, Germany
Borax is a naturally occurring mineral compound that is commonly used as a laboratory reagent. It consists of sodium, boron, and oxygen, and its primary function is to serve as a buffer and source of borate ions in various laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Boric acid, by Merck Group (8 mentions)
Sodium hydroxide (naoh), by Merck Group (6 mentions)
Hydrochloric acid (hcl), by Merck Group (6 mentions)
Toluidine blue, by Merck Group (4 mentions)
Zwittergent 3 14 detergent, by Merck Group (4 mentions)
3 hydroxydiphenol, by Merck Group (4 mentions)
Sodium chloride (nacl), by Merck Group (3 mentions)
Glucuronic acid, by Merck Group (3 mentions)
Poly l lysine (pll), by Merck Group (2 mentions)
Chloroform, by Merck Group (2 mentions)
H2so4, by Merck Group (2 mentions)
Hexane, by Merck Group (2 mentions)
Sodium carbonate, by Merck Group (2 mentions)
Hydrogen peroxide, by Merck Group (2 mentions)
Axiophot, by Zeiss (2 mentions)
Tween 20, by Merck Group (2 mentions)
Bovine serum albumin (bsa), by Merck Group (2 mentions)
Vector shield fluorescence mounting medium, by Vector Laboratories (2 mentions)
Potassium chloride (kcl), by Merck Group (2 mentions)
Alizarin red, by Merck Group (2 mentions)
44 protocols using borax
1
Primary Cortical Cell Culture Preparation
2
Quantification of Glutathione Metabolites
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GSH, GSSG, γ-Glu-Cys, Cys-Gly, 5-oxo-proline, Glu and the other amino acids used in the study, CH3OH, CD3OD (99.8 atom % 2H), acetone, pentafluoropropionic anhydride, 2,3,4,5,6-pentafluorobenzyl bromide, ophthalmic acid (γ-glutamyl-α-amino-n-butyryl-glycine) and borax (for the preparation of borate buffer) were purchased from Sigma-Aldrich (Steinheim, Germany). Hydrochloric acid (ultrapure, 37%) was from AppliChem (Darmstadt, Germany). Stock solutions were prepared in, and diluted with, deionized water, as appropriate. Glassware for GC-MS (1.8-mL autosampler vials and 0.2-mL microvials) were purchased from Macherey-Nagel (Düren, Germany).
Safety Considerations. PFPA is corrosive and malodorous. PFB-Br is corrosive and a lachrymator. Inhalation and contact with skin and eyes should be avoided. All work should be, and was, performed in a well-ventilated fume hood.
Safety Considerations. PFPA is corrosive and malodorous. PFB-Br is corrosive and a lachrymator. Inhalation and contact with skin and eyes should be avoided. All work should be, and was, performed in a well-ventilated fume hood.
3
Valorization of Agricultural and Industrial Wastes
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The camote and cassava peels were collected from a local vendor of native delicacies, and the sawdust utilized for particle board production was collected from the shavings of a local furniture shop in Vigan City, Philippines. The sawdust is typically from the shavings of mahogany, gmelina, and bamboo varieties. Borax (Sigma-Aldrich, Darmstadt, Germany), 10 wt. % sodium hypochlorite (Sigma-Aldrich, Darmstadt, Germany), 98 wt. % sodium hydroxide, hydrochloric acid (Sigma-Aldrich, Darmstadt, Germany), and 37 wt. % formaldehyde (Sigma-Aldrich, Darmstadt, Germany) were used. All methods were carried out in three trials.
4
Histological Analysis of Mouse Brain
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Mouse brains were fixed in 4% PFA and were subjected to paraffin embedding and sectioning. Serial 5 µM sections were used for conventional H&E staining, immunohistochemistry staining or immunofluorescence as previously described [60 ]. For immunohistochemistry staining, sections were incubated with cleaved-caspase3 primary antibody. For BrdU staining, tissue sections were denatured with 0.1 N HCl for 1 h in a 37 °C water bath. After denaturation, the sections were neutralized with 0.1 M Borax at pH 8.5 (Sigma) for 10 min. Sections were washed with 0.3% Triton X-100/1×PBS (wash buffer) 3 times and blocked with 5% normal donkey serum (Sigma-Aldrich) in wash buffer for 1 h at room temperature. All immunofluorescence-labeled images were acquired using a Nikon C2 + confocal microscope.
5
Transwell Invasion Assay for Renal Cancer
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The invasiveness of renal cancer cells was assessed by examining the invasion of 24 wells through Matrigel‐coated Transwell inserts with 8 μm pores (BD Biosciences) according to the manufacturer's protocol. Cells (2.0 × 105) in 500 mL serum‐free MEM were added to each insert, and 500 mL of MEM containing 10% fetal bovine serum with or without 5 μmol/L vorinostat and/or 10 μmol/L fluvastatin was added to the bottom of each well. Forty‐eight hours later, the cells that had remained inside the inserts were removed and cells that had migrated through the inserts' membranes were fixed in methanol and stained with 1% toluidine blue (Kanto Chemicals) in 1% borax (Sigma). The cells were counted in 3 randomly chosen visual fields at ×200 magnification.
6
BrdU Incorporation Assay for Cell Proliferation
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Cells grown in the presence of 33 μM BrdU for the indicated time were fixed with 75% ethanol (Panreac), permeabilized with 2N HCl (Panreac) for 30 min at room temperature and pH equilibrated using 0.1 M BORAX (Sigma) for 2 min. Cells were incubated with a mouse anti-BrdU antibody (BD) in 1% BSA, 1 × PBS for 1 h at room temperature, washed and stained with a donkey anti-mouse FITC-conjugated antibody (Jackson). Stained cells were treated with RNase A (Sigma) followed by DNA staining with 2.5 μg μl–1 propidium iodide (Sigma) overnight at 4 °C. BrdU intensities were acquired by FACS Calibur and analysed using the FlowJo software. Antibody details are given in Supplementary Table 2 .
7
Magnesia-Based Cementitious Paste Formulation
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MPC pastes were prepared by blending various powders, including magnesia (magnesium oxide) (MgO > 99%,Merck KGaA Frankfurter Str.25, Darmstadt, Germany), ADP (NH4H2PO4 > 99%, Sigma-Aldrich (Darmstadt, Germany)), borax, which was used as a set retarder, and aluminum. All these materials were sourced from Sigma Aldrich. Magnesia was subjected to calcination at 1500 °C for 6 h to reduce its reactivity. Different quantities of renfort powder were added to the mixture. Aggregates were deliberately excluded from this research to avoid any additional interference from their impurities.
8
Fabrication of AM1241-Loaded PEG-DTT Hydrogels
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PEG-DTT hydrogels were prepared by the Michael addition reaction using poly(ethylene glycol) diacrylate (PEGDA) (molecular weight: 700, Sigma-Aldrich, United States) and DTT (Sigma-Aldrich, United States) over the sodium tetraborate catalyst. AM1241 (0, 1, 2.5, 5, 10, 20, 40, 80, 120, and 200 µM) (Sigma-Aldrich, United States) was then immediately added to this solution and mixed to prepare a finished AM1241-loaded PEG-DTT hydrogel. First, 100 mg of PEGDA (0.14 mM) and 22 mg of DTT (0.14 mM) were dissolved in 0.5 ml of water. Then, 0.5 ml of 0.1 mol/L borax (Sigma-Aldrich, United States) solution was added to the mixture and stirred vigorously for 10 s. Keep the solution at room temperature (25°C) for a while and check the gelation time by tube inversion.
9
Toluidine Blue Staining for Cell Morphology
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The influence of 1d and 1h was evaluated on cells density and morphology by toluidine-blue staining. A total of 2 × 104 cells/well were cultured in 24-well plates and were incubated at 37 °C and 5% CO2 with tested compounds at 5 µM. After 48 and 72 h, adherent cells were fixed with 70% cold ethanol and stained with 1% toluidine blue and 1% borax (Sigma Aldrich, St. Louis, MO, USA). Cells were then observed by an inverted microscope connected with a camera at 40×, 100×, and 200× magnifications (Leica, Wild Heer-brugg, Wetzlar, Germany).
10
Biochemical Assays for Natural Compounds
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Hexane, chloroform, ethanol, sodium carbonate, NBT, EDTA, hydroxylamine hydrochloride, hydrogen peroxide, sulphosalicylic acid, DTNB, KCl, ferric chloride, HCl, TCA, TBA, BHT, naphthylethylene diamine dihydrochloride, sodium nitroprusside, sulphanilic acid, NaOH, H2SO4, para-dimethyl amino-benzaldehyde, n-Propanol, Na2CO3, Borax, carbazole etc. were purchased from Sigma–Aldrich, Merck and Himedia. All the chemicals and solvents used were of analytical grade.
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