Lactulose, a probe used to check paracellular permeability, was added at 40 mM/ml final concentration to the apical side of all monolayers at time 0. Samples were collected from the basolateral side at increasing time intervals (ranging from 30 min to 6 h) after gliadin and L.GG treatments. Lactulose concentration was measured by high performance anion exchange chromatography (HPAEC) [22 (link)]. After deproteination with acetonitrile 1:1 v/v, samples were centrifuged at 4000 rpm for 10 min, the supernatant collected, filtered through a 0.22 mm membrane (Millipore, Bedford, Mass., USA), and diluted with water 1 to 10 (basolateral samples) or 1 to 100 (apical samples). HPAEC coupled with pulsed amperometric detection (HPAEC-PAD) was performed on a Dionex Model ICS-5000 with a gold working electrode and a 25 μl peek sample loop (Dionex Corp., Sunnyvale, CA, USA). Carbohydrate separation was carried out by a Carbopac PA-10 pellicular anion-exchange resin connected to a Carbopac PA-10 guard column at 30°C.
0.22 mm membrane
Manufactured by Merck Group
Sourced in United States
The 0.22 mm membrane is a laboratory filtration device designed to remove small particles and microorganisms from liquids. It has a pore size of 0.22 mm, which allows the passage of fluids while effectively retaining a wide range of contaminants. This membrane is commonly used in various scientific and industrial applications to purify, sterilize, or clarify solutions.
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Lab products found in correlation
Basic fibroblast growth factor (bfgf), by Lonza (1 mentions)
Vascular endothelial growth factor (vegf), by Lonza (1 mentions)
Heparin, by Lonza (1 mentions)
Insulin like growth factor, by Lonza (1 mentions)
Fetal bovine serum (fbs), by Lonza (1 mentions)
Amphotericin b, by Lonza (1 mentions)
Gentamicin, by Lonza (1 mentions)
Hydrocortisone, by Lonza (1 mentions)
Ascorbic acid, by Lonza (1 mentions)
Human epidermal growth factor (hegf), by Lonza (1 mentions)
2 protocols using 0.22 mm membrane
1
Quantifying Paracellular Permeability via Lactulose
2
Endothelial Cell Inflammation Modeling
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Human aortic endothelial cell line (HAEC) (Lonza), a human aortic endothelial primary cell obtained at passage 2. These cells are very well characterised and were cultured in Endothelial Cell Basal Medium-two and the following growth supplements: hydrocortisone, basic fibroblast growth factor, vascular endothelial growth factor, insulin-like growth factor, ascorbic acid, heparin, FBS, human epidermal growth factor, gentamicin, amphotericin B (Lonza). Experiments were performed at passage 4 where native LDL and Mox-LDL (100 mg/ml), IL-18 (250 pg/ml) and TNF-a (10 ng/ml), sterilised by filtration through a 0.22-mm membrane (Millipore), were added to the culture medium and incubated with the endothelial cells for 24 hours at 37 C under 5% CO 2 . The experiments were performed in triplicate.
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