HDL3-signalling in HepG2 cells involves glycosyl-phospatidylinositol-anchored proteins

Nazih‐Sanderson, Françoise; Pinchon, Gaelle; Nion, Stéphane; Fruchart, Jean‐Charles; Delbart, Christiane

doi:10.1016/s0005-2760(97)00017-9

Cited by 10 publications

(2 citation statements)

References 43 publications

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“…2. Add the mixes at a 1:1 ratio to cells prepared as above, i. e. at a final cell density of 10x10 6 cells/ml and a final MBCD concentration of 2.5 mM.…”

Section: Establishing Extraction Conditions Where No Net Cholesterol mentioning

confidence: 99%

“…Cellular cholesterol depletion can be achieved in several ways; treatment with metabolic inhibitors, growing cells in lipoprotein deficient medium or using chemicals to immobilize or to extract cholesterol. Examples of these methods are inhibition of cholesterol biosynthesis by statins [4], the culture of cells in the absence of exogenous cholesterol [5], the use of cholesterol binding agents such as digitonin, filipin and saponin [6,7] and oxidation of cholesterol [8]. The most commonly used method is acute cholesterol depletion using MBCD.…”

Section: Introductionmentioning

confidence: 99%

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Cholesterol Depletion Using Methyl-β-cyclodextrin

Mahammad

Parmryd

2014

Methods in Molecular Biology

220

166

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Cholesterol is an essential component of mammalian cells. It is the major lipid constituent of the plasma membrane and is also abundant in most other organelle membranes. In the plasma membrane cholesterol plays critical physical roles in the maintenance of membrane fluidity and membrane permeability. It is also important for membrane trafficking, cell signalling and lipid as well as protein sorting. Cholesterol is essential for the formation of liquid ordered domains in model membranes, which in cells are known as lipid nanodomains or lipid rafts. Cholesterol depletion is widely used to study the role of cholesterol in cellular processes and can be performed over days using inhibitors of its synthesis or acutely over minutes using chemical reagents. Acute cholesterol depletion by methyl-β-cyclodextrin (MBCD) is the most widely used method and here we describe how it should be performed to avoid the common side-effect cell death.

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“…2. Add the mixes at a 1:1 ratio to cells prepared as above, i. e. at a final cell density of 10x10 6 cells/ml and a final MBCD concentration of 2.5 mM.…”

Section: Establishing Extraction Conditions Where No Net Cholesterol mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Cholesterol Depletion Using Methyl-β-cyclodextrin

Mahammad

Parmryd

2014

Methods in Molecular Biology

220

166

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Phospholipids of Plasma Lipoproteins, Red Blood Cells and Atheroma, Analysis of

Ravandi

Kuksis

2000

Encyclopedia of Analytical Chemistry

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The large number of lipid classes and great complexity of molecular species present in blood plasma, red cells, platelets and atheromatous lesions requires a combination of analytical techniques for comprehensive analyses, including chemical and enzymatic derivatization of samples. In many instances only partial analyses are required, which can be accomplished by specific analytical techniques. This chapter describes the full spectrum of the methodology ranging from the most basic thin‐layer chromatography (TLC) to the detailed mass spectrometric assays. Traditional sample extraction by liquid–liquid partition is time‐consuming and involves large volumes of solvents. Liquid–solid extraction using adsorbent cartridges is more economical. At the present time total lipid extracts can be effectively assayed for lipid class content and molecular species composition by flow injection tandem mass spectrometry (MS/MS), while a more detailed analysis of complex lipid mixtures is provided by a combination of liquid chromatography with on‐line electrospray mass spectrometry (LC/ESMS). The new techniques permit completion of the analyses in a few hours, where previously several days or weeks may have been required. The soft ionization mass spectrometric techniques have permitted the recognition and detailed analysis of such minor components of blood as the lipid oxidation and glycation products often observed in disease.

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