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4 protocols using 2 4 2 hydroxyethyl piperazin 1 yl ethanesulfonic acid

1

Preparation of High-Purity Aqueous Solutions

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N-[tris(hydroxymethyl)methyl]glycine (Tricine, purity > 99 wt%), 2-[(2-hydroxy-1,1-bis(hydroxymethyl)ethyl)amino]ethane sulfonic acid (TES, purity > 99 wt%), 2-(N-morpholino) ethanesulfonic acid (MES, purity > 99 wt%), 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (HEPES, purity > 99.5 wt%), 2-(cyclohexylamino)ethanesulfonic acid (CHES, purity > 99 wt%), and [P4444][OH] (45 wt% in water), were purchased from Sigma–Aldrich. Sodium hydroxide pellets were supplied from Eka Chemicals. BSA (fraction V) was obtained from Acros Organics. Methanol (HPLC grade, purity > 99.9 wt%) was obtained from Fisher Scientific (UK). Acetonitrile (purity > 99.7 wt%) was supplied from Lab-Scan (Ireland). Purified water passed through a reverse osmosis and a Milli-Q plus 185 water purifying system was used in all experiments. Sodium nitrate (purity > 99.5 wt%) was obtained from Himedia Labs. Potassium nitrate (purity > 98.0), nitric acid (65 wt%), and potassium hydrogen phthalate (purity > 99.8) were purchased from Panreac (Barcelona, Spain).
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2

Radiolabeling and Characterization of MSCs

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8‐Hydroxyquinoline (oxine), sodium citrate, NaOAc, Na2CO3, and 2‐[4‐(2‐hydroxyethyl)‐piperazin‐1‐yl] ethane sulfonic acid (HEPES) solution were purchased from Sigma–Aldrich (St Louis, MO). Dimethyl sulfoxide (DMSO) was obtained from Acros Organics (Belgium, USA). DFS was purchased from Innochem (Beijing, China). DFP was purchased from MedChem Express Ltd. Deferoxamine mesylate (DFO) was purchased from Novartis China. Gallium nitrate hydrate and zirconium tetrachloride were obtained from Innochem (Beijing, China). 89Zr‐oxalate was obtained from Dongcheng AMS Pharmaceutical Co., Ltd (Nanjing, China). 68Ga3+ solution was eluted from a 68Ge/68Ga generator (ITG, Germany) with 4 mL of 0.05 M HCl (Merck, Germany), from which the 0.5 mL fraction with the highest radioactivity was used for further experiments. β‐Actin internal reference primers, MSCs markers primer sequences used for real‐time qPCR were obtained from Sangon Biotech Co., Ltd. (Shanghai, China).
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3

Proteomic Analysis Reagents and Suppliers

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2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid, phenylmethanesulfonylfluoride (PMSF), 2.5-dihydroxybenzoic acid, ethylenediaminetetraacetic acid (EDTA), nicotinamide adenine dinucleotide phosphate, sodium dithionite, trypsin, and sodium deoxycholate were purchased from Sigma-Aldrich (St. Louis, MO, USA); acetonitrile and trifluoroacetic acid were purchased from ICN Pharmaceuticals Inc. (Costa Mesa, CA, USA); Coomassie Brilliant Blue R-250 was purchased from Fluka (Seelze, Germany). Modified trypsin (catalog no. V511C) was obtained from Promega (Madison, WI, USA). Other chemicals were purchased from Reakhim-Penza, LLC (Penza, Russia).
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4

Heavy Metal Ion Detection via DNA Aptamers

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Tris-(8-hydroxyquinoline)aluminum (Alq3, 99.995%), 2-morpholinoethanesulfonic acid (MES, 99%) and 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (HEPES, 99%) were purchased from Sigma-Aldrich. Tetrahydrofuran (THF, 99%) was purchased from Aladdin. ssDNA and Tris-HCl buffer solution were purchased from Shanghai Shenggong Biological, China. Sequences of ssDNA which can specifically bind with four heavy metal ions are as follows:

Pb2+: 5′-GGTTGGTGTGGTTGG-3′;

Hg2+: 5′-TTCTTTCTTCCCCTTGTTTGTT-3′;

Cd2+: 5′-GGGTTCACAGTCCGTT-3′;

As3+: 5′-ATGCAAACCCTTAAGAAAGTGGTCGTCCAAAAAACCATTG-3′.

Nitric acid (HNO3, 65.0~68.0%) was purchased from Sinopharm Chemical Reagent. Shrub branches and leaves were obtained from the Institute of Geophysical and Geophysical Exploration, Chinese Academy of Geological Sciences.
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