Ethylene diamine tetraacetic acid (edta)

Manufactured by Mallinckrodt

Sourced in United States

EDTA is a chemical compound used in laboratory applications as a chelating agent. It forms stable complexes with metal ions, which can be useful in various analytical and purification processes. EDTA's core function is to bind and sequester metal ions, making it a versatile tool for laboratory work.

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5 protocols using ethylene diamine tetraacetic acid (edta)

Synthesis and Characterization of Pyridyl Porphyrins

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meso-Tetrakis(2-N-pyridyl)porphyrin (H₂T-2-PyP) and meso-tetrakis(3-N-pyridyl)porphyrin (H₂T-3-PyP) were purchased from Frontier
Scientific. Ethyl p-toluenesulfonate (98%) was from
Sigma-Aldrich. The n-octyl p-toluenesulfonate
and methyl-tri-n-octylammonium chloride (>95%) were from TCI America.
MnCl₂·4H₂O (99.7%) was supplied by J. T.
Baker, FeCl₂ (98%) was from Sigma-Aldrich, and NH₄PF₆ (99.99%) was from GFS chemicals. Anhydrous diethyl
ether and acetone were from EMD chemicals, while dichloromethane,
chloroform, acetonitrile, EDTA, and KNO₃ were purchased
from Mallinckrodt. Anhydrous N,N-dimethylformamide
(DMF) of 99.8% purity (kept over 4-Å molecular sieves) and plastic-backed
silica gel TLC plates (Z122777-25EA) were from Sigma-Aldrich. Xanthine,
equine ferricytochrome c (lot 7752), and (+)-sodium l-ascorbate (>98%) were from Sigma, whereas xanthine oxidase
was prepared by R. Wiley.^{1 (link)} Triethylamine
(Et₃N) of >99.5% purity was obtained from Thermo Scientific
Pierce. All chemicals were used as received without further purification.
The ¹H NMR spectra were recorded on a spectrometer “Mercury
Varian 300” with deuterated chloroform as solvent.

Tovmasyan A., Carballal S., Ghazaryan R., Melikyan L., Weitner T., Maia C.G., Reboucas J.S., Radi R., Spasojevic I., Benov L, & Batinic-Haberle I. (2014). Rational Design of Superoxide Dismutase (SOD) Mimics: The Evaluation of the Therapeutic Potential of New Cationic Mn Porphyrins with Linear and Cyclic Substituents. Inorganic Chemistry, 53(21), 11467-11483.

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Exosome Protein Analysis by Western Blot

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Cells and exosomes were lysed in a RIPA buffer containing 1% NP-40, 20 mM Tris-HCl (pH 7.5), 137 mM NaCl, 0.5 mM EDTA (Mallinckrodt Baker Inc., Philipsburg, NJ, USA), 10% glycerol (Frutarom LTD, Haifa, Israel), 1% protease inhibitor cocktail (Sigma-Aldrich) and 0.1% SDS (Biological Industries). After centrifugation, protein content was quantified using the Bradford assay, and 25 μg of protein from each specimen were heat-denaturated in a Laemmli sample buffer and fractionated in 10% gels by SDS–PAGE, electro-transferred to nitrocellulose membrane for blotting with antibodies. In order to block nonspecific binding, membranes were incubated for 1 h in 5% low-fat milk dissolved in TBST. Membranes containing proteins originating from exosomes did not undergo blocking as per the manufacturer’s instruction for CD81 antibody. Membranes were then incubated with the following antibodies: ERK monoclonal antibody (#4695, Cell Signaling Biotechnology, Danvers, MA, USA), p-ERK monoclonal antibody (#4377, Cell Signaling Biotechnology) and CD81 polyclonal antibody (PA5-13582, Thermo Scientific Inc., Waltham, MA, USA). GAPDH (14C10; Cell Signaling Biotechnology) was used as loading control. Blots were developed in Bio-Rad ChemiDoc™ XR and analyzed using Image Lab™ software.

Onallah H., Mannully S.T., Davidson B, & Reich R. (2022). Exosome Secretion and Epithelial-Mesenchymal Transition in Ovarian Cancer Are Regulated by Phospholipase D. International Journal of Molecular Sciences, 23(21), 13286.

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Preparation of Pyridinium-Based Compounds

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H₂T-2-PyP, H₂T-3-PyP and H₂T-4-PyP were purchased from Frontier Scientifi, 2-Methoxyethyl tosylate (>98%), 4-methoxybuthanol (>98%), 6-bromohexan-1-ol (>95%), from TCI America, 5-methoxypenthanol (98%) from Karl Industries Inc., p-toluenesulfonyl chloride (98%) from Alpha Aesar, pyridine (99%) and tetra-n-butylammonium chloride hydrate (98%) from Aldrich, MnCl₂ × 4H₂O (99.7%) and hexane from J. T. Baker and NH₄PF₆ (99.99% pure) from GFS chemicals. Anhydrous diethyl ether and acetone were from EMD chemicals; absolute methanol, ethyl acetate (EtOAc), dichloromethane, chloroform, acetonitrile, EDTA and KNO₃ were from Mallinckrodt; 98% anhydrous N,N-dimethylformamide (DMF, kept over 4-Å molecular sieves), plastic-backed silica gel thin layer chromatography (TLC) plates (Z122777-25EA) were from Sigma-Aldrich; and silica (SiliaFlash® G60, 70-230 mesh) was from Silicycle (Canada). 6-Methoxyhexan-1-ol and H₂TMOE-2-PyP⁴⁺ were prepared as previously described.,^{26 (link), 27} All other chemicals were used as received.

Rajic Z., Tovmasyan A., de Santana O.L., Peixoto I.N., Spasojevic I., do Monte S.A., Ventura E., Rebouças J.S, & Batinic-Haberle I. (2017). Challenges encountered during development of Mn porphyrin-based, potent redox-active drug and superoxide dismutase mimic, MnTnBuOE-2-PyP5+, and its alkoxyalkyl analogues. Journal of inorganic biochemistry, 169, 50-60.

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Antioxidant Activity Assay Protocol

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α,α-Diphenyl-β-picrylhydrazyl free radical (DPPH), linoleic acid, butylated hydroxyanisole (BHA), butylated hydroxytoluene, α-tocopherol, bovine serum albumin, thiobarbituric acid, ferrozine, lecithin, SDS (sodium dodecyl sulfate), ammonium thiocyanate, ferric chloride, KH₂PO₄, and K₂HPO₄ were purchased from Sigma-Aldrich (St. Louis, MO, USA). Sodium dihydrogen phosphate, disodium hydrogen phosphate, NaBr, and trichloacetic acid were obtained from Merck & Co. Inc. (Kenilworth, JN, USA). Tween 20 was obtained from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). HCl, NaCl, and copper sulfate were purchased from the Tokyo Chemical Industry Co. Ltd. (Tokyo, Japan). EDTA was purchased from Mallinckrodt Pharmaceuticals (Raleigh, NC, USA). Ferrous chloride, Coomassie brilliant blue G-250, n-butanol, and phosphotungstic acid were bought from Avantor Performance Materials (Baker analyzed reagents; Center Valley, PA, USA).

Lin Y.P., Lin L.Y., Yeh H.Y., Chuang C.H., Tseng S.W, & Yen Y.H. (2016). Antihyperlipidemic activity of Allium chinense bulbs. Journal of Food and Drug Analysis, 24(3), 516-526.

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Histological Analysis of Pregnant Pubic Joint

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PS and IpL were fixed with 4% paraformaldehyde (Merck, Darmstadt, Germany) in 0.1 M phosphate-buffered saline (PBS; pH 7.4) for 24 hours at 4°C. Decalcification was performed during five days in 5% ethylenediaminetetraacetic acid (EDTA, Mallinckrodt Baker, Phillipsburg, NJ, USA) and 2% paraformaldehyde in 0.1 M PBS, pH 7.4 for five days at 4°C. Tissues of three animals per experimental group (a total of 18 animals) were dehydrated in graded concentrations of alcohol, embedded in Historesin (Leica Microsystems, Heidelberg, Germany) and sectioned (3 μm) for staining with Giemsa [14 (link)]. Alternatively, interpubic tissues of an additional three animals per group (a total of 18 animals) were decalcified and dehydrated in graded concentrations of alcohol, embedded in paraffin, sectioned (5 μm), stained with Sirius Red F3B and observed with the aid of polarization microscopy to gain insight into the time-dependent changes of the microstructure of collagen fibres in the pubic joint during pregnancy [12 (link)]. Sections were examined and imaged under a Nikon Eclipse E800 light microscope (Nikon Corporation, Tokyo, Japan).

Castelucci B.G., Consonni S.R., Rosa V.S., Sensiate L.A., Delatti P.C., Alvares L.E, & Joazeiro P.P. (2018). Time-dependent regulation of morphological changes and cartilage differentiation markers in the mouse pubic symphysis during pregnancy and postpartum recovery. PLoS ONE, 13(4), e0195304.

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