Platelet-rich plasma (PRP) was obtained by collecting blood in acid citrate dextrose solution (citric acid 130 mM, Trisodic citrate 152 mM, and glucose 112 mM). Samples were then diluted in Tyrode solution (HEPES 10 mM, NaCl 137 mM, KCl 2.9 mM, NaHCO3 12 mM; pH 7.4) and centrifuged at 200g for 7 minutes with deceleration at 27°C. To isolate platelets, 0.02 U/mL apyrase (A6237), 1 μM of prostaglandin E1 (P5515), and 10 μM of indomethacin (I7378-5G) provided by Sigma Aldrich, were added to PRP before being recentrifuged at 850g for 6 minutes at 27°C and suspended in Tyrode. Platelets were counted as previously described.19 (link)
Prostaglandin e1 p5515
Manufactured by Merck Group
Prostaglandin E1 (P5515) is a laboratory reagent produced by Merck Group. It is a naturally occurring lipid compound that functions as a potent vasodilator and regulator of various physiological processes. The core function of this product is to serve as a research tool for studying the role of prostaglandin E1 in biological systems.
Automatically generated - may contain errors
Lab products found in correlation
Naproxen, by Merck Group (1 mentions)
Ketoprofen, by Merck Group (1 mentions)
Ortho phosphoric acid, by Scharlab (1 mentions)
Lc ms grade methanol, by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
Ibuprofen, by Merck Group (1 mentions)
Diclofenac, by Merck Group (1 mentions)
Vanillin, by Merck Group (1 mentions)
Chloroform, by Merck Group (1 mentions)
Hplc grade acetonitrile and water, by Merck Group (1 mentions)
2 protocols using prostaglandin e1 p5515
1
Platelet-rich Plasma Isolation Protocol
2
Quantifying NSAID Metabolite Profiles
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The NSAIDs diclofenac (sodium salt, D6899, DIC), ketoprofen (≥ 98% TLC, K1751, KET), naproxen (≥ 98%, sodium salt, M1275, NAP) and Ibuprofen (sodium salt, ≥ 98% GC, I1892, IBU) were purchased from Sigma-Aldrich. Stocks solutions of 4 mg ml -1 for each NSAIDs were prepared in Methanol (LC-MS grade, Merck). For the confirmation of the metabolites, Prostaglandin E1 (P5515) and Arachidonic acid (23401) standards from Sigma Aldrich were used. Lipid extraction was based on the method of Zöllner and Kirsch (1962) , using Chloroform (HPLC grade, Merck), Methanol (HPLC grade, Sigma Aldrich) for the extraction and for the analysis Vanillin (99%, V1104, Sigma Aldrich) and orthophosphoric acid (85%, reagent grade, Scharlau) were used. The mobile phase used for metabolomics approach was HPLC grade acetonitrile and water purchased from Merck (Darmstadt, Germany).
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