Dulbecco’s Modified Eagle Medium (DMEM), fetal bovine serum (FBS), and trypLE Express were purchased from Life Technologies (Carlsbad, CA, USA). CellTiter 96® AQueous One Solution was purchased from Promega (Madison, WI, USA). ELISA BrdU (colorimetric) was acquired from Roche (Basel, Switzerland). BD BioCoatTM MatrigelTM Invasion Chambers were acquired from Corning® (Corning, New York, NY, USA). The dead cell apoptosis kit with Annexin V-FITC/ PI was obtained from BD Bioscience (Franklin Lakes, NJ, USA). RNAse A and PI were purchased from Invitrogen (Carlsbad, CA, USA). Anti-caspase 8 and anti-caspase 9 antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). IRDye 800CW Goat anti-mouse IgG and IRDye 800CW Goat anti-rabbit IgG were acquired from LI-COR (Lincoln, NE, USA). Anti-caspase 3 was purchased from Abcam (Cambridge, UK). L-α-phosphatidylcholine (PC) and anti-vinculin antibody were obtained from Sigma-Aldrich (St. Louis, MI, USA). 1,2-Dioleoyl-sn-glycerol-3-phosphoserine sodium salt (Lipoid PS 18:1/18:1) was obtained from Lipoid GmbH (Ludwigshafen, Germany) and LabAssay Phospholipid was obtained from Wako (Osaka, Japan).
Labassay phospholipid
Manufactured by Fujifilm
Sourced in Germany
The LabAssay Phospholipid is a lab equipment product from Fujifilm. It is designed to measure the concentration of phospholipids in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Labassay cholesterol, by Fujifilm (2 mentions)
Anti caspase 3, by Abcam (1 mentions)
Anti caspase 8, by Santa Cruz Biotechnology (1 mentions)
Anti caspase 9, by Santa Cruz Biotechnology (1 mentions)
L α phosphatidylcholine pc, by Merck Group (1 mentions)
Irdye 800cw goat anti mouse igg, by LI COR (1 mentions)
Irdye 800cw goat anti rabbit igg, by LI COR (1 mentions)
Brdu elisa, by Roche (1 mentions)
Anti vinculin antibody, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Tryple express, by Thermo Fisher Scientific (1 mentions)
Rnase a, by Thermo Fisher Scientific (1 mentions)
Celltiter 96 aqueous one solution, by Promega (1 mentions)
Dulbecco s modified eagle medium dmem, by Thermo Fisher Scientific (1 mentions)
Amplex red cholesterol assay, by Thermo Fisher Scientific (1 mentions)
Free cholesterol e, by Fujifilm (1 mentions)
Dc protein assay kit 2, by Bio-Rad (1 mentions)
Hdl cholesterol e, by Fujifilm (1 mentions)
Labassay triglyceride, by Fujifilm (1 mentions)
7 protocols using labassay phospholipid
1
Comprehensive Cell Viability and Apoptosis Assay
2
Quantification of Cellular Cholesterol and Phospholipids
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One million cells were plated in 150‐mm petri dishes 24 h before the experiment. Cells were treated with 10 mM methyl‐β‐cyclodextrin (MβCD, C4555 Sigma) for 30 min for the positive control. Cells were washed three times and scraped in 500 μl of PBS. Cells in PBS (500 μl) were mixed with 2 ml of EtOH/chloroform (2/1) solution in glass tubes. After 5 min of centrifugation at 1,400 g, the supernatant was transferred into a new tube. Then, 0.5 ml of 50 mM citric acid, 1 ml of water, and 0.5 ml of chloroform were added. After 20 min of centrifugation at 1,400 g, the lower phase‐containing lipids were transferred in new glass tubes and dried in a stream of nitrogen. For the lipids quantification, dried lipids were solubilized in EtOH 95% solution. Cholesterol quantification was performed with the Amplex Red cholesterol assay (A12216 Invitrogen), and phospholipids quantification was performed with LabAssay phospholipid (296‐63801 Wako Chemicals) according to the manufacturer instructions.
3
Dietary Phospholipid Modulation of Brain
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The experiment was conducted using the same schedule as that in Experiment 1. Two LPC-containing food additives, LIPOID R LPC 20 and SLP-LPC70, which were distributed as enzyme-modified lecithin, were used in addition to GPC. The total Ch content (free plus [lyso]phospholipid forms) of the test food additives was quantified using LabAssay ™ Phospholipid (Wako Pure Chemical Industries, Ltd.). The test food additives and GPC were orally administered to the rats at 100 mg Ch equivalent/kg BW (2000, 1280, and 246 mg/kg BW for LIPOID R LPC 20, SLP-LPC70, and GPC, respectively). Following the same SCO treatment and fixation procedure as that in Experiment 1, the brains were harvested and divided into the frontal cortex, hippocampus, and striatum.
4
Comprehensive Protein and Lipid Analysis
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Protein concentrations were measured using DC Protein Assay Kit II (Bio-Rad). Phospholipid and total and free cholesterol levels were evaluated enzymatically (LabAssay Phospholipid, LabAssay Cholesterol, and Free Cholesterol E; Wako Diagnostics). Preβ1-HDL was measured using ELISA kit (Sekisui/American Diagnostica GmbH) according to the manufacturer’s recommendations.
Protein conjugate analysis by size exclusion chromatography coupled to multiple angle light scattering was performed as described in theData Supplement . Data collection and analysis were performed in the ASTRA software using ASTRA’s Protein Conjugate Analysis method.32 The differential refractive index increments (dn/dc) of 0.185, 0.150, and 0.130 were used for molar mass determination of apo AI, phosphatidylcholine and mixture of mammalian lipids, respectively.33 (link),34 (link)
Protein conjugate analysis by size exclusion chromatography coupled to multiple angle light scattering was performed as described in the
5
Quantifying Bovine MFGM Proteins and Rotavirus
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Two MFGM-enriched commercial products, high-fat WPC and BMP, were purchased from Fonterra Cooperative Group Ltd. Protein content was measured using the Kjeldahl method (ISO, 2014). Phospholipid content was measured using LabAssay Phospholipid (Takayama et al., 1977; (link)Wako Chemicals) according to the manufacturer's instructions. Bovine lactadherin content was measured using the FLEXIQuant method as described below. The human rotavirus MO strain was kindly provided by Osamu Nakagomi and Toyoko Nakagomi (Department of Molecular Microbiology and Immunology, Nagasaki University, Nagasaki, Japan), and a virus titer was determined by the focus forming assay according to a previously described (Ebina et al., 1990) (link).
Because no human or animal subjects were used, this analysis did not require approval by an Institutional Animal Care and Use Committee or Institutional Review Board.
Because no human or animal subjects were used, this analysis did not require approval by an Institutional Animal Care and Use Committee or Institutional Review Board.
6
Enzymatic Phospholipid Quantification
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Choline-containing phospholipid content of NL and NE were determined using an enzymatic kit (LabAssay™ Phospholipids, Fujifilm Wako Chemicals, Neuss, Germany) according to manufacturer’s instructions.
7
Lipid Profiling in Treated Mice
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After treatment for 8 weeks, all mice were euthanized with pentobarbital sodium injection. Serum and the other major tissues were collected and stored at −80 °C for further analysis.
The extraction of lipids in liver was performed with chloroform-methanol (2:1, v/v) according to the Folch’s method. Subsequently, the concentrations of triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL), and phospholipids (PL) in liver and serum were determined using the kits LabAssay™ Triglyceride, LabAssay™ Cholesterol, HDL-Cholesterol-E, and LabAssay™ Phospholipids obtained from FUJIFILM Wako (Tokyo, Japan), respectively. All experiments were performed based on the manufacturer’s protocols.
The extraction of lipids in liver was performed with chloroform-methanol (2:1, v/v) according to the Folch’s method. Subsequently, the concentrations of triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL), and phospholipids (PL) in liver and serum were determined using the kits LabAssay™ Triglyceride, LabAssay™ Cholesterol, HDL-Cholesterol-E, and LabAssay™ Phospholipids obtained from FUJIFILM Wako (Tokyo, Japan), respectively. All experiments were performed based on the manufacturer’s protocols.
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