Ethylenediaminetetraacetic acid edta

The 10, 40 and 147 kDa dextrans (Catalog number: 1179876, 1179865 and D-487); bovine serum albumin (BSA); sodium chloride; ferrozine; butylated hydroxyanisole (BHA); pyrocatechol violet; and ascorbic acid were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Potassium ferricyanide, ferrous sulfate (II), ethylenediaminetetraacetic acid (EDTA), Gallic acid, ammonium molybdate, hydrogen peroxide 30%, acetic acid, Folin–Ciocalteu phenol reagent, ethanol and sulfuric acid were obtained from Merck (Darmstadt, Germany). Culture media components Minimum essential Dulbecco’s modified Eagle medium (DMEM), l-Glutamine, sodium pyruvate, sodium bicarbonate, non-essential amino acids, fetal bovine serum and phosphate buffered saline (PBS) were acquired from Invitrogen Corporation (Burlington, ON, Canada). All other reagents and solvents were of analytical degree.

Brains were dissected following anesthesia and trans-cardiac perfusion with ice-cold saline, tissue samples were snap-frozen in liquid N₂ and stored at − 80 °C. Cortices were homogenized using a Potter-Elvejhem homogenizer (20 strokes, 700 rpm; VWR, Leuven, Belgium) in 6 vol of ice-cold Tris-proteinase-phosphatase-inhibitor buffer, containing 25 mM Tris–HCl (pH 8.1), 150 mM sodium chloride (Sigma), 1 mM ethylene diamine tetra-acetic acid (EDTA, Merck), 1 mM ethylene glycol tetra-acetic acid (EGTA, Sigma-Aldrich), 5 mM sodium pyrophosphate (Sigma), 5 mM sodium fluoride (Sigma-Aldrich), 1 mM PMSF (Sigma), 1 mM sodium vanadate (Sigma-Aldrich), and a cocktail of proteinase inhibitors (Roche) and phosphatase inhibitors (Roche).

Human antithrombin (ATIII)
(R1), purified human thrombin (R2), and chromogenic substrate specific
for thrombin (CS-01(38)) (R3) were purchased from Hyphen-Biomed (France).
European Pharmacopoeia reference standard was purchased from EP (Strasbourg
Cedex, France). Tris(hydroxymethyl)aminomethane, polyethylene glycol
6000, hydrochloric acid, and acetic acid were purchased from Sigma-Aldrich
(USA). Ethylenediaminetetraacetic acid (EDTA) and sodium chloride
(NaCl) were purchased from Merck (Germany). All materials detailed
above were of analytical grade.

Carnosine was purchased from Sigma (St. Louis, MO, USA). 4,2-Hydroxyethyl,1-piperazineethanesulfonic acid (HEPES), 3-(N-morpholino) propane sulfonic acid (MOPS), Dimethyl sulfoxide (DMSO), D-mannitol, bovine serum albumin (BSA), thiobarbituric acid (TBA), 3-[4,5dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT), Coomassie brilliant blue, Rhodamine 123 (Rh 123), Ethylene glycol-bis (2-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), Sodium succinate, Hydroxymethyl aminomethane hydrochloride (Tris-HCl), and ethylenediaminetetraacetic acid (EDTA) were purchased from Merck (Darmstadt, Germany). All salts for preparing buffer solutions (analytical grade) were purchased from Merck (Darmstadt, Germany).

The polymer PEICT with an average molecular weight of 46,800 (g/mol) was supplied by SK Chemicals, Gyeonggi-do, Korea [8 (link)]. Chloroform (99%), and trifluoroacetic acid (TFA) (98%) were purchased from Wako, Pure Chemical Industries Ltd., Osaka, Japan. Ethanol (99.8%) was obtained by Merck, Japan. Cell migration on PEICT ENMs was performed using NIH 3T3 mouse fibroblast obtained from ATCC (Manassas, VA), USA [24 (link)]. The 24-well cell culture flasks and Petri-dishes were obtained from SPL Life Sciences (Pocheon, Korea). Ethylenediaminetetraacetic acid (EDTA) was purchased from Merck (KGaA), Germany. Dulbecco’s modified Eagle’s medium (DMEM) was supplied by Nacalai Tesque, Kyoto, Japan. Fetal bovine serum (FBS) and Dulbecco’s phosphate-buffered saline (DPBS) were provided by HyClone (Logan, UT, USA). The Cell Counting Kit-8 (CCK-8) reagent was obtained from Dojindo Laboratories Japan [25 (link)].

Murashige and Skoog’s (MS) basal medium, agar-agar, and diluent solution for DNA were obtained from Hi-media, India. Bavestien^® (Carbendazimpowder) was procured from ASF India Limited, New Delhi, India. Tween-20 (Polysorbate 20), mercury (II) chloride, lead (II) nitrate, methanol, n-hexane, acetonitrile, acetone, ethyl acetate, diethyl ether, nitric acid (HNO₃), perchloric acid (HClO₄), sucrose, sodium hydroxide (NaOH), ethylenediaminetetraacetic acid (EDTA) and phytagel were obtained from Merck, Merck-Sigma Aldrich, St. Louis, MO, USA. All standard phenolic compounds viz., gallic acid, caffeic acid, p-coumaric acid, ellagic acid, rosmarinic acid, trans-cinnamic acid and vanillic acid were procured from Sigma Aldrich, Merck-Sigma Aldrich, St. Louis, MO, USA. All the solvents used in these experiments were HPLC grade.

5,5′-dithiols-(2-nitrobenzoic acid) (DTNB), 2,4,6-tris (2-pyridyl) -s-triazine (TPTZ), and thiobarbituric acid (TBA) were obtained from Sigma Chemical Co (St Louis, MO, USA). Hydrochloric acid (HCL), trichloroacetic acid (TCA), formaldehyde, ethylenediaminetetraacetic acid (EDTA) and 2, 4-dinitrophenylhydrazine (DNPH) were gained from Merck (Germany). Fluvoxamine was purchased from Abidi Pharmaceutical Co (Tehran, Iran).