The freeze-dried supernatants were reconstituted by dissolving in 100 μL of filtered 3 mM perchloric acid and analyzed by HPLC using an LC-2000Plus system (JASCO, Tokyo, Japan) equipped with two RSpak KC-811 columns (Showa Denko, Tokyo, Japan). Quantification of organic acids was achieved using a solution containing 0.2 mM bromothymol blue in 15 mM sodium phosphate buffer, with peak detection occurring at 445 nm. The column was maintained at a temperature of 60°C, and the flow rates were 0.7 mL/min for the 9 mM perchloric acid and 1.2 mL/min for the 0.2 mM bromothymol blue solution. To calibrate the measurements, standard powders of succinate, malate, fumarate, lactate, and acetate were employed, all of which were obtained from Fujifilm Wako Chemicals. Citrate powders were purchased from Nacalai Tesque, INC. (Kyoto, Japan).
Lactate
Manufactured by Fujifilm
Sourced in Japan, United States
Lactate is a laboratory equipment product designed to measure the concentration of lactate in various biological samples. It functions by detecting and quantifying the levels of lactate, a metabolite produced during anaerobic respiration. This equipment provides accurate and reliable data to support various clinical and research applications.
Automatically generated - may contain errors
Lab products found in correlation
Fumarate, by Fujifilm (1 mentions)
Lc 2000plus system, by Jasco (1 mentions)
Rspak kc 811 column, by Resonac (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Fujifilm (1 mentions)
Sodium hydrate, by Fujifilm (1 mentions)
Chir99021, by Selleck Chemicals (1 mentions)
Mtesr1, by Merck Group (1 mentions)
Matrigel, by Merck Group (1 mentions)
Y 27632, by Merck Group (1 mentions)
Matrigel, by Corning (1 mentions)
Mtesr1, by STEMCELL (1 mentions)
H9 hesc, by WiCell (1 mentions)
3 protocols using lactate
1
Quantification of Organic Acids by HPLC
2
Porcine Blood pH Adjustment and Stimulation
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Lactate (Wako, Osaka, Japan) or sodium hydrate (Wako) was added to the priming solution to adjust the pH of porcine blood to approximately 6.8 or 8.0. The pH values of porcine blood were analyzed with Epoc (Epocal Inc., Ottawa, Canada). For adjusting the blood temperature, the reservoir was warmed up or cooled down with a water bath. In some experiments, porcine blood was stimulated with 0.6 µg/mL phorbol 12-miristate 13-acetate (PMA) (Wako) at 37 °C for 120 minutes before adding to the circuit. For inhibiting endogenous DNase activity, EDTA (Wako) was added to the priming solution at a concentration of 5 mM.
3
Cardiac Differentiation of Human Embryonic Stem Cells
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Human embryonic stem cell (hESC)-derived cardiac differentiation was induced as previously described [9 (link)]. For cardiac lineage induction, H9 hESCs (WiCell, Madison, WI, USA) were plated onto Matrigel (Corning, New York, NY, USA)-coated 6 well plate at a density of 0.5 × 106 cells/well and cultured in mTeSR1 (STEMCELL Technologies, Vancouver, BC, Canada) supplemented with cGMP for 2–3 days. Three days before cardiomyocyte differentiation, hESCs were plated on Matrigel-coated 6 well plate at a density of 0.6 × 106 cells/well and cultured in mTeSR1 supplemented with 5 μM Y27632 (Sigma-Aldrich, St. Louis, MO, USA). The culture medium was replaced with RPMI+B27 medium (RPMI1640 with B27 supplement without insulin; Gibco, Grand Island, NY, USA) supplemented with CHIR99021 (10 μM, S1263; Selleckchem, Houston, TX, USA) for 3 days. The culture medium was subsequently replaced with RPMI+B27 supplemented with IWP2 (5 μM; Tocris, Bristol, UK), followed by culture for 5 days. On day 8, the culture medium was replaced with RPMI+B27. The medium was changed every 1–2 days. Beating cardiomyocytes were observed on days 9–10. Cardiomyocytes were chemically sorted by lactate (Wako, Richmond, VA, USA).
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