Alpha-glucosidase enzyme from Saccharomyces cerevisiae, α-amylase from porcine pancreas and 2-chloro-4-nitrophenyl-α-D-maltotrioside (CNPG3) were purchased from Sigma Aldrich. All other chemicals were purchased from UFC Biotechnology. A microplate reader (Anthos Zenyth-200RT, Cambridge, UK) was used for the spectrophotometric measurements. A Bruker NMR Spectrometer (Bruker, Yokohama, Japan) used for 1H-NMR (400 MHZ) and 13C-NMR (100 MHz) analysis. Spectra were recorded in CD3OD or DMSO-d6 using tetramethylsilane (Sigma-Aldrich, St. Louis, MO, USA) as an internal standard, and chemical shift values were expressed in δ ppm.
α amylase from porcine pancreas
Manufactured by Merck Group
Sourced in United States, Germany
α-amylase from porcine pancreas is an enzyme that catalyzes the hydrolysis of starch into simpler sugars. It is a commonly used laboratory reagent for the analysis and characterization of starch and other carbohydrates.
Automatically generated - may contain errors
Lab products found in correlation
α glucosidase from saccharomyces cerevisiae, by Merck Group (10 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (8 mentions)
Acarbose, by Merck Group (7 mentions)
Gallic acid, by Merck Group (7 mentions)
Pancreatin from porcine pancreas, by Merck Group (7 mentions)
3 5 dinitrosalicylic acid, by Merck Group (6 mentions)
Pepsin from porcine gastric mucosa, by Merck Group (6 mentions)
Ascorbic acid, by Merck Group (6 mentions)
Folin ciocalteu reagent, by Merck Group (5 mentions)
P nitrophenyl α d glucopyranoside, by Merck Group (4 mentions)
Orlistat, by Merck Group (4 mentions)
Methanol, by Merck Group (4 mentions)
Porcine bile extract, by Merck Group (4 mentions)
2 2 azino bis 3 ethylbenzothiazoline 6 sulfonic acid, by Merck Group (3 mentions)
Lipase from porcine pancreas, by Merck Group (3 mentions)
L ascorbic acid, by Merck Group (3 mentions)
Acetylthiocholine iodide, by Merck Group (3 mentions)
Acetonitrile, by Thermo Fisher Scientific (3 mentions)
Amyloglucosidase from aspergillus niger, by Merck Group (3 mentions)
Quercetin, by Merck Group (3 mentions)
48 protocols using α amylase from porcine pancreas
1
Enzymatic Assays for Saccharomyces cerevisiae
2
Measuring α-Amylase Activity in Tea
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α-Amylase from porcine pancreas (Sigma-Aldrich) was dissolved in PBS at a concentration of 0.06 mg/mL. Tea extracts were mixed 15:100 with the assay buffer from the MaxDiscovery™ Amylase Assay Kit (Bioo Scientific, Texas, USA). The α-amylase activity of the mixture was measured according to the manufacturer’s instructions.
3
Phytochemical Extraction and Bioassays
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Analytical grade solvents for extraction and chromatography were purchased from Echo Chemical Co. (Taiwan). α-Glucosidase from Bacillus stearothermophilus, α-amylase from porcine pancreas, 3,5-dinitrosalicylic acid (DNS), 4-nitrophenyl phosphate disodium salt hexahydrate (pNPP), 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical, 3-(2-pyridyl)-5,6-diphenyl-1,2,4-triazine-4′,4″-disulfonic acid sodium salt (Ferrozine), trizma hydrochloride (Tris–HCl), DL-dithiothreitol (DTT), (+)-catechin hydrate, acarbose and rutin hydrate were purchased from Sigma Chemical Co. (USA). p-Nitrophenyl-α-d -glucopyranoside (pNPG), KNaC4H4O6·4H2O, quercetin dihydrate, ethylenediaminetetraacetic acid disodium salt (EDTA-Na2), FeCl2·4H2O, NH4Fe(SO4)2·12H2O and thiazolyl blue tetrazolium bromide (MTT) were purchased from Acros (Belgium). Ursolic acid, citric acid, vanillin and AlCl3·6H2O were purchased from Merck (Germany). PTP1B from human was purchased from Enzo Life Sciences (Switzerland). Dulbecco's modified Eagle's medium (DMEM) and newborn calf serum (NCS) were purchased from Gibco BRL (USA). Melacacidin was separated and purified from Acacia confusa root according to Lin and Chang.18 All other unlabelled chemicals and reagents were purchased from Sigma Chemical Co. (USA).
4
Inhibition Assay for α-Amylase
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The enzyme inhibition activity against α-amylase from porcine pancreas (Sigma-Aldrich, USA) were determined using 1 mM of 2-chloro-4 nitrophenol-α-D-maltotroside-3 as substrate, following a method as described previously by Yonemoto et al.[15 (link)] with modifications. In 96 well plate, 30 μl of enzyme solution (25 U/ml) and 30 μl of tested inhibitors in DMSO were mixed and preincubated at 37°C for 10 min. Then, 30 μl of substrate were added and incubated again at 37°C for 20 min. The absorbance was measured at 405 nm using microplate reader. All tests were analyzed in triplicate. The percent inhibition was calculated as aforementioned formula.
5
Enzymatic Inhibition of α-Amylase Activity
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Inhibition of α-amylase activity was assayed using the enzymatic assay and reagents proposed by Sigma Aldrich. Briefly, the pre-incubation mix was composed of 1.5 mL of buffer (50 mmol/L sodium phosphate, 50 mmol/L sodium chloride, 0.5 mmol/L calcium chloride, pH 6.9), 0.4 mL of the solutions to be tested containing the inhibitors at different concentrations, and 0.1 mL of the enzymatic solution consisting of 100 U/mL of α-amylase from porcine pancreas (Sigma Aldrich). It was pre-incubated for 30 min at 25°C. Then, 0.5 mL of this pre-mix was incubated with 0.5 mL of a substrate composed of 1% (w:v) gelatinized potato starch (Sigma Aldrich) in 20 mmol/L sodium phosphate buffer and 6.7 mmol/L sodium chloride (pH 6.9) for 20 min at 25°C. The reaction was stopped by the addition of 0.5 mL of a solution composed of sodium potassium tartrate and 96 mmol/L 3,5-dinitrosalicylic acid, then by boiling for 15 min, then by cooling on ice. The absorbance was read at 540 nm. The assays were conducted in triplicates. Acarbose (Sigma Aldrich) was used as a positive control.
6
Enzymatic Analysis of Amylase and Antioxidants
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α-Amylase from porcine pancreas (EC 3.2.1.1), 2,2’-Azinobis 3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 2,2’-diazobis-(2-aminodinopropane)-dihydrochloride (AAPH), 2,2-diphenyl-1-picrylhydrazyl (DPPH), Folin–Ciocalteu (FC) reagent, gallic acid (GA), 6-hydroxy-2,5,7,8-tetramethyl-2-carboxylic acid (Trolox), and sodium carboxymethylcellulose (CMC) were obtained from Sigma-Aldrich, Co. (St. Louis, MO, USA). Amyloglucosidase (EC 3.2.1.3), glucose oxidase-peroxidase (GOPOD) and 1.3:1.4 mixed-linkage β-glucan kits were provided by Megazyme International Ireland (Wicklow, Ireland).
7
Antioxidant and Enzyme Inhibition Assays
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Acetonitrile, formic acid, methanol, ABTS (2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid), 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), 2,4,6-tri(2-pyridyl)-s-triazine (TPTZ), 2,2-Di(4-tert-octylphenyl)-1-picrylhydrazyl (DPPH), methanol, acetic acid, phosphate buffered saline (pH 7.4), LOX Activity Assay kit, AChE Assay kit, α-amylase from porcine pancreas, α-glucoamylase from Rhizopus sp., lipase from porcine pancreas, trini-trobenzenesulfonic acid, NaH2PO4, and 3,5-dini-trosalicylic acid were purchased from Sigma-Aldrich (Steinheim, Germany). COX Inhibitor Screening Assay Kit was purchased from Cayman (No. 560131; Ann Arbor, MI, USA). (−)-Epicatechin, (+)-catechin, chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, dicaffeic acid, procyanidin A2, procyanidin B2, p-coumaric acid, caffeic acid, 4-caffeoylquinic, kampferol-3-O-galactoside, quercetin-3-O-rutinoside, quercetin-3-O-galactoside, quercetin-3-O-glucoside, cyanidin-3-O-arabinoside, cyanidin-3-O-xyloside, cyanidin-3-O-galactoside, and cyanidin-3-O-glucoside were purchased from Extrasynthese (Lyon, France). Acetonitrile for ultra-performance liquid chromatography (UPLC; gradient grade) and ascorbic acid were from Merck (Darmstadt, Germany). The carriers (30%) applied to produce powders were maltodextrin DE (20–40) and inulin (Beneo-Orafti, Belgium).
8
Colorimetric Assay for Porcine Amylase
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The α-amylase from porcine pancreas was purchased from Sigma. Hano et al. [32 (link)] described chromogenic method was employed to evaluate soluble α-amylase activity. In brief, phosphate buffer (0.1 M, pH6.8) was used to prepare enzyme at 1u/mL concentration and thoroughly mixed in 5mM of 4-nitrophenyl-α-D-maltopentaoside (s4NPM; Sigma). After half hour incubation at room temperature, solution was column filtered to stop reaction and sodium carbonate (1M) solution was added in equal volume. Increase in absorbance value compared to blank at 405 nm was used to determine enzymatic activity. The % enzyme inhibition for each sample was calculated by subtraction of absorbance values in the presence and absence of calli extracts.
9
Jaboticaba By-product Valorization
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The jaboticaba (Myrciaria cauliflora) processing by-product, consisting of peel, seed, and remaining pulp (10 kg) was acquired from Sítio do Bello (Piraibuna, SP, Brazil). DES components such as malic acid were purchased from Êxodo Científica (Sumaré, SP), 2-Hydroxyethyltrimethylammonium chloride (Choline Chloride – ChCl) >98% purity was obtained from Sigma-Aldrich (Steinheim, Germany) and propylene glycol (>99.5) from Neon Commercial (Suzano, SP, Brazil). Solvents such as 99.5% ethanol, 98–100% formic acid, and ethyl acetate 99.5%, from Êxodo Científica (Sumaré, SP, Brazil). Other chemicals as Amberlite XAD-7HD resin, methanol suitable for HPLC (≥99,9% purity), TPTZ (2,4,6-tri(2-pyridyl)-s-triazine), ABTS (2,2′-azinobis-3-ethylbenzothiazoline-6-sulfonic acid), pNPG (p-nitrophenyl-α-d -glucopyranoside), pNPB (p-nitrophenol-butyrate), DNS (3,5-dinitro salicylic acid), standards Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) (97% purity) and gallic acid (>97% purity), and enzymes α-amylase from porcine pancreas, α -glucosidase from Saccharomyces cerevisiae and lipase from human pancreas were acquired from Sigma Aldrich (Steinheim, Germany).
10
Formulation and In Vitro Digestion of Blackcurrant Extract
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The ingredients used to prepare the model systems were dried blackcurrant (Ribes nigrum) pomace (BCP), supplied by the Institute of Natural Materials Technology (Technische Universität Dresden, Germany) and prepared by drying the fresh pomace at 70 °C for 2 h and milling it in a ZM 100 ultracentrifuge mill (Retsch GmbH, Haan, Germany) at 14,000 rpm using a 1 mm sieve [9 (link)] (Reißner et al., 2019). The blackcurrant extract (BCE) was prepared from BCP, olive oil (Consum, Valencia, Spain), native wheat starch (C*Gel, Cargill BV, Amsterdam, Netherlands), and whey protein isolate from milk (Harrison Sport Nutrition S.L, Granada, Spain). All the enzymes (α-amylase from porcine pancreas, pepsin from porcine gastric mucosa, porcine bile extract, lipase from porcine pancreas, and pancreatin from porcine pancreas) used in the in vitro digestion analysis were supplied by Sigma-Aldrich (Barcelona, Spain).
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