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Ace from porcine kidney

Manufactured by Merck Group
Sourced in Italy

ACE from porcine kidney is a laboratory reagent used for various research and analytical applications. It is an enzyme that plays a role in the regulation of blood pressure. The core function of this product is to serve as a tool for researchers and scientists in their investigations, without any specific interpretation or extrapolation on its intended use.

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5 protocols using ace from porcine kidney

1

Casein Hydrolysate Bioactive Peptides

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Dulbecco’s modified Eagle’s medium (DMEM), L-glutamine, fetal bovine serum (FBS), phosphate buffered saline (PBS), penicillin/streptomycin and 96-well plates were purchased from Euroclone (Milan, Italy). Acetonitrile (ACN), formic acid, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), sitagliptin, Gly-Pro-amido-4-methylcoumarin hydrobromide (Gly-Pro-AMC) and ACE from porcine kidney were from Sigma-Aldrich (St. Louis, MO, United States). ACE1 Activity Assay Kits come from Biovision (Milpitas Blvd., Milpitias, CA, United States). The DPP-IV inhibitor screening assay kit was bought from Cayman Chemical (Michigan, United States). Caco-2 cells were obtained from INSERM (Paris, France). Casein hydrolysates (CH), which is spray dried samples from the production process, was supplied by A. Costantino S.R.L. (Italy). (Casein hydrolysate sport/health batch 218F0004, 100 g). Casein hydrolysates (CH), which is spray dried samples from the production process, was supplied by A. Costantino S.R.L. (Italy). (Casein hydrolysate sport/health batch 218F0004, 100 g).
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2

ACE-inhibitory activity of peptides

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The ACE-inhibitory activity of the CH was evaluated following previously reported methods (51 (link), 52 (link)) and tested by measuring with HPLC the formation of hippuric acid (HA) from hippuryl-histidyl-leucine (HHL), used as mimic substrate for ACE. Tests were performed in 100 mM Tris-HCOOH, 300 mM NaCl pH 8.3 buffer, and using ACE from porcine kidney (Sigma-Aldrich, Milan, Italy). CH hydrolysate was tested at 0.08, 0.17, 0.35, 0.7 and 1.0 mg/mL.
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3

ACE Inhibitory Activity Assay

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ACE inhibitory activity was proven by measuring, with HPLC, the development of hippuric acid (HA) using hippuryl-histidyl-leucine (HHL) as a mimic substrate for ACE I. FPH was tested at 1 mg/mL in 100 mM Tris-HCOOH, 300 mM NaCl pH 8.3 buffer, and using ACE from porcine kidney (Sigma-Aldrich, Milan, Italy). All experimental details of sample preparations and analyses conditions have been published elsewhere [30 (link),31 (link)].
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4

ACE Inhibitory Activity Assay

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ACE inhibitory activity was tested by measuring with HPLC the formation of hippuric acid from hippuryl-histidyl-leucine, used as a mimic substrate for ACE I. Test was performed in 100 mM Tris-HCOOH, 300 mM NaCl pH 8.3 buffer, and using ACE from porcine kidney (Sigma-Aldrich, Milan, Italy). S, M and T peptides mixtures were tested at 1.0 mg/mL and IC50 value is the concentration needed to observe a 50% inhibition of ACE activity. All experimental details of samples preparation and analyses conditions have been published elsewhere (Boschin et al., 2014a) (link) (Boschin et al., 2014b) (link).
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5

Casein Purification and Characterization

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Casein (Promilk 85) was obtained from Ingredia (Arrax Cedex, France). ACE from porcine kidney, bicinchoninic acid protein assay kit, Hammarsten casein and Triton X-100 were purchased from Sigma (St. Louis, MO, USA).
Glucose was obtained from Panreac (Barcelona, Spain), bacteriological peptone was purchased from Cultimed (Barcelona, Spain) and yeast extract and agar were acquired from Pronadisa (Madrid, Spain). The fluorogenic substrate o-aminobenzoyl-Gly-p-nitro-Phe-Pro was provided by Bachem Feinchemikalien (Bubendorf, Switzerland). Coomassie Brilliant Blue G-250, 30% acrylamide/bis solution 29:1 and Precision Plus Protein Dual Color standards were obtained from Bio-Rad Laboratories (Hercules, CA, USA).
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