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3 protocols using sodium carbonate

1

Apoptosis Assay in Cell Cultures

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Dulbecco’s Modified Eagle’s Medium (DMEM) and all cell culture reagents were purchased from Life Technologies (CA, USA). Fetal bovine serum (FBS) and MTT were obtained from Himedia. FITC AnnexinV Apoptosis Detection Kit was bought from Becton Dickinson (NJ, USA). All other chemicals were of analytical grade. A colorimetric assay kit was used to measure caspase-3 activity (R&D Systems, Inc., Minneapolis, MN, USA). Enzyme-linked immunonosorbent assay (ELISA) kits were used to measure Bax and Bcl-2 protein levels (Elabscience, Houston, TX, USA). Ethanol solution (60%), for extraction procedures, was prepared using ethanol PA from Synth (Brazil). Sodium carbonate and aluminum chloride were obtained from Dinâmica (Brazil) and Synth, respectively, while Folin-Ciocalteu was obtained from Merck®. Gallic acid and quercetin standards were purchased from Sigma (Annapolis, MD, USA). Methanol was used for UPLC/MS procedures, acetonitrile and formic acid were HPLC grade from Merck®, and sodium formate was prepared from sodium hydroxide (Dinâmica, Brazil) and formic acid (Merck®).
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2

Synthesis of Inorganic Compounds

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The
chemicals magnesium
chloride hexahydrate (MgCl2·6H2O, >99%),
aluminum chloride hexahydrate (AlCl3·6H2O, >99%), and sodium carbonate (NaCO3, >99.5%) were
purchased
from Dinâmica and used as received. Sodium hydroxide (NaOH,
>97%) was purchased from Synth. All experimental solutions were
prepared
using deionized water, which was also used to rinse and clean the
samples.
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3

Guava Leaves Antioxidant Potential

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The leaves of the guava plant (Psidium guajava Linn. var. Pomifera) from the seedling bed was kindly donated by the Division of Parks and Conservation Units of the Secretariat for the Environment (Sorocaba, São Paulo, Brazil), ethanol (Synth, São Paulo, Brazil), hydrochloric acid (Neon, São Paulo, Brazil). Folin-Ciocalteau reagent (Dinâmica, São Paulo, Brazil), sodium carbonate (Dinâmica, São Paulo, Brazil), gallic acid (Dinâmica, São Paulo, Brazil). 2,2-diphenyl-1-picrylhydrazyl (Sigma-Aldrich, Steinheim, Germany), methanol (Chemco, São Paulo, Brazil). All reagents used were analytical grade.
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