methyl β cyclodextrin  (Millipore)


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    Structured Review

    Millipore methyl β cyclodextrin
    Methyl β Cyclodextrin, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/methyl β cyclodextrin/product/Millipore
    Average 99 stars, based on 36 article reviews
    Price from $9.99 to $1999.99
    methyl β cyclodextrin - by Bioz Stars, 2020-02
    99/100 stars

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    Related Articles

    Conditioned Place Preference:

    Article Title: Epigallocatechin-3-gallate reduces inflammation induced by calcium pyrophosphate crystals in vitro
    Article Snippet: Cells were then treated with CPP (final concentration 0.025 mg/ml; InvivoGen) for 24 h in presence or absence of EGCG (10, 50 μM) and 2% FCS. .. In some experiments, before stimulation with crystals FLS and THP-1 were pretreated for 1 h with EGCG at the indicated concentrations or for 45 min with 10 mM MβCD (Sigma-Aldrich) prepared from a 500 ng/ml PBS mother solution.

    Cell Isolation:

    Article Title: Cytotoxicity of purified listeriolysin O on mouse and human leukocytes and leukaemia cells
    Article Snippet: Paragraph title: Eukaryotic cell isolation and growth conditions ... For some experiments cell cultures were supplemented with the following compounds to modify the LLO activity: cholesterol (Sigma), NCS containing 150 μg/ml cholesterol (390 μM), and MβCD (Sigma).

    Cytometry:

    Article Title: Mechanistic Insights into the Cholesterol-dependent Binding of Perfringolysin O-based Probes and Cell Membranes
    Article Snippet: Paragraph title: Flow cytometry ... Cells were washed with PBS and then incubated at 37 °C for 30 min in 0.5 ml of PBS containing 5 mM mβCD (Sigma) complexed with the indicated concentration of cholesterol.

    Construct:

    Article Title: Dynamic lateral organization of opioid receptors (kappa, muwt and muN40D) in the plasma membrane at the nanoscale level, et al. Dynamic lateral organization of opioid receptors (kappa, muwt and muN40D) in the plasma membrane at the nanoscale level
    Article Snippet: Approximately, 24 to 36 hours (GPI and VSVG constructs) and 48 hours (opioid receptors) after transfection, cells were fixed as described before. .. Forty‐eight hours posttransfection, cells were incubated with 10 mM MβCD (Sigma) in full DMEM supplemented with 10 mM HEPES and 1 mg/mL BSA for 30 minutes at 37°C, similarly as described before.

    Article Title: GPI-anchored receptor clusters transiently recruit Lyn and G? for temporary cluster immobilization and Lyn activation: single-molecule tracking study 1
    Article Snippet: To examine the involvement of Lyn in STALL induction, SYF cells were transfected with the cDNA for Lyn fused with GFP at the C terminus (Lyn-GFP); this GFP construct is known to be functional ( ). .. Partial depletion of cholesterol in the plasma membrane was performed by incubating the cells in 4 mM MβCD (Sigma-Aldrich) at 37°C for 30 min ( ) or in 60 μg/ml saponin (Sigma-Aldrich) on ice for 15 min ( ).

    Incubation:

    Article Title: Uptake of Helicobacter pylori Vesicles Is Facilitated by Clathrin-Dependent and Clathrin-Independent Endocytic Pathways
    Article Snippet: Chemical inhibitors were added to AGS cells prior to vesicle incubation. .. MβCD (Sigma-Aldrich) in F-12K (+FBS) was added at 4 mM or 10 mM for 15 min before vesicle addition both with and without MβCD present.

    Article Title: Telocinobufagin and Marinobufagin Produce Different Effects in LLC-PK1 Cells: A Case of Functional Selectivity of Bufadienolides
    Article Snippet: .. In 24-well plates, 1–2 × 104 LLC-PK1 cells/well were treated with 1, 10 or 100 nM CTS for 24, 48, and 72 h. In order to investigate the involvement of intracellular signaling pathways, 100 nM CTS were incubated with or without inhibitors of Src (SU6656, 10 μM; Sigma-Aldrich, St. Louis, MO, USA), ERK1/2-MEK1/2 (U0126, 10 μM; Cell Signaling Technology, Danvers, MA, USA), PI3K ( , 5 μM; Cell Signaling Technology, Danvers, MA, USA), p38 (SB202190, 10 μM; Cell Signaling Technology, Danvers, MA, USA), JNK1/2 (SP600125, 1.5 μM; Cell Signaling Technology, Danvers, MA, USA), or GSK-3β (BIO, 0.5, 1, and 5 μM; Sigma-Aldrich, St. Louis, MO, USA) for 72 h. For MβCD (Sigma-Aldrich, St. Louis, MO, USA) assay, cells were pretreated with 10 mM MβCD for 30 min and then 1 mM MβCD + 100 nM TCB in 2.5% FBS for 72 h. At these time points, two wells from each group were trypsinized and the number of Trypan blue-viable cells was counted in Neubauer chamber (hemocytometer). .. In 96-well plates, 3 × 103 LLC-PK1 cells/well were treated with 100 nM CTS with or without inhibitors of Src (SU6656, 10 μM), ERK1/2-MEK1/2 (U0126, 10 μM), PI3K ( 5 μM), p38 (SB202190, 10 μM), JNK1/2 (SP600125, 1.5 μM), or GSK-3β (BIO, 0.5, 1 and 5 μM) for 72 h. MTT was added directly to culture wells and incubated for 4 h. The absorbance was measured at 570 nm with a 96-well plate reader.

    Article Title: Distinct molecular and cellular contributions to stabilizing selectin-mediated rolling under flow
    Article Snippet: .. Alternatively, streptavidin-coated cells were incubated with 10 mM MβCD or αCD (Sigma-Aldrich) in 100 μl HBSS for 15 min at 37°C and then diluted into 1 ml HBSS with 0.5% human serum albumin. .. In other experiments, streptavidin-coated cells were incubated with 10 μM cytochalasin D (Sigma-Aldrich) or control DMSO diluent for 20 min at 37°C.

    Article Title: Dynamic lateral organization of opioid receptors (kappa, muwt and muN40D) in the plasma membrane at the nanoscale level, et al. Dynamic lateral organization of opioid receptors (kappa, muwt and muN40D) in the plasma membrane at the nanoscale level
    Article Snippet: .. Forty‐eight hours posttransfection, cells were incubated with 10 mM MβCD (Sigma) in full DMEM supplemented with 10 mM HEPES and 1 mg/mL BSA for 30 minutes at 37°C, similarly as described before. ..

    Article Title: Mechanistic Insights into the Cholesterol-dependent Binding of Perfringolysin O-based Probes and Cell Membranes
    Article Snippet: .. Cells were washed with PBS and then incubated at 37 °C for 30 min in 0.5 ml of PBS containing 5 mM mβCD (Sigma) complexed with the indicated concentration of cholesterol. .. The cells were then washed twice with PBS and incubated at 4 °C for 20 min with 0.5 μM of the indicated PFO derivative (10% labeled with Alexa488) in 100 μl of PBS.

    Article Title: Cholesterol modulates open probability and desensitization of NMDA receptors
    Article Snippet: .. For acute cholesterol depletion by MβCD, DIV 6–7 CGCs were exposed to low-potassium medium supplemented with 5 m m MβCD (mean molecular weight 1310 g mol−1 ; Aldrich, St Louis, MO, USA) and incubated at 37°C in 5% CO2 for 1–60 min as indicated. .. Chronic cholesterol depletion by simvastatin was carried out by incubating DIV 5 CGCs in low-potassium medium supplemented with 100 n m simvastatin (Sigma) for 4 days prior to patch-clamp measurements (Zacco et al .).

    Article Title: A role for myosin-1A in the localization of a brush border disaccharidase
    Article Snippet: .. For studies involving cholesterol depletion, cells were incubated in 2% MβCD in serum-free DME (Sigma-Aldrich) for 30 min before fractionation. .. Cloning of GFP-M1A-tail and SINT-YFP fusion constructs The GFP-M1A-tail and SINT-YFP fusion constructs were assembled using standard molecular biological techniques.

    Article Title: Binding of Human Immunodeficiency Virus Type 1 to Immature Dendritic Cells Can Occur Independently of DC-SIGN and Mannose Binding C-Type Lectin Receptors via a Cholesterol-Dependent Pathway
    Article Snippet: .. As controls for the effective functioning of amantadine and MβCD, drug-treated DC (as described above) were incubated with fluorescein isothiocyanate (FITC)-conjugated transferrin substrate (50 μg/ml) (Sigma) for 1 h at 4°C and then shifted to 37°C for 1 h, cytospun onto coverslips, and then fixed in 4% paraformaldehyde. .. Coverslips were rinsed extensively with PBS, stained with DAPI (4′,6′-diamidino-2-phenylindole; 0.2 μg/ml in distilled H2 O) for 5 min, washed once with distilled H2 O, dehydrated in 100% methanol for 15 min, air dried, mounted in Vectashield (Vector Laboratories, Burlingame, Calif.), and sealed with nail polish.

    Activity Assay:

    Article Title: Cytotoxicity of purified listeriolysin O on mouse and human leukocytes and leukaemia cells
    Article Snippet: .. For some experiments cell cultures were supplemented with the following compounds to modify the LLO activity: cholesterol (Sigma), NCS containing 150 μg/ml cholesterol (390 μM), and MβCD (Sigma). .. The affinity chromatography of LLO Ten ml of E. coli suspension (approximately 108 /ml) grown in the presence of 1 mM IPTG was centrifuged at 4400 × g for 10 min at 4°C, and the supernatant was removed.

    Expressing:

    Article Title: A β-Glucan-Based Dietary Fiber Reduces Mast Cell-Induced Hyperpermeability in Ileum From Patients With Crohn’s Disease and Control Subjects
    Article Snippet: To study uptake mechanisms of β-glucan, two endocytosis inhibitors were used; MβCD, which chelates cholesterol from the plasma membrane, has previously been shown to decrease the translocation of β-glucan expressing fungi by inhibiting lipid raft formation. .. Upon M cell-transformation, monolayers were washed and preincubated with HBSS with or without the addition of 3 mM MβCD (Sigma-Aldrich) or 30 µM CPZ (Fluka) at 37°C for 20 minutes.

    Modification:

    Article Title: Dynamic lateral organization of opioid receptors (kappa, muwt and muN40D) in the plasma membrane at the nanoscale level, et al. Dynamic lateral organization of opioid receptors (kappa, muwt and muN40D) in the plasma membrane at the nanoscale level
    Article Snippet: COS‐7, PANC‐1, MCF‐7 and MDA‐MB‐468 cells were cultured in phenol red‐free Dulbecco's modified eagle medium (DMEM) supplemented with 10% FBS, 1 mM sodium pyruvate, 100 units/mL penicillin, 100 units/mL streptomycin and 2 mM l ‐alanyl‐l ‐glutamine (full DMEM). .. Forty‐eight hours posttransfection, cells were incubated with 10 mM MβCD (Sigma) in full DMEM supplemented with 10 mM HEPES and 1 mg/mL BSA for 30 minutes at 37°C, similarly as described before.

    Over Expression:

    Article Title: Exosomal miR-9 released from HIV Tat stimulated astrocytes mediates microglial migration
    Article Snippet: Chemical inhibitors, including MβCD and DMA were purchased from Sigma (St. Louis, MO, USA). .. The miR-9 overexpression plasmid has-miR-9-1, miR-control, and miR-9 inhibitor plasmid hsa-miR-9-5p-locker/hsa-miR-9-3p-locker as well as miR-locker-control plasmids and lentiviruses were purchased from Biosettia (San Diego, CA).

    Translocation Assay:

    Article Title: A β-Glucan-Based Dietary Fiber Reduces Mast Cell-Induced Hyperpermeability in Ileum From Patients With Crohn’s Disease and Control Subjects
    Article Snippet: Paragraph title: Studies of Yeast-derived β-glucan Translocation ... Upon M cell-transformation, monolayers were washed and preincubated with HBSS with or without the addition of 3 mM MβCD (Sigma-Aldrich) or 30 µM CPZ (Fluka) at 37°C for 20 minutes.

    Flow Cytometry:

    Article Title: Mechanistic Insights into the Cholesterol-dependent Binding of Perfringolysin O-based Probes and Cell Membranes
    Article Snippet: Paragraph title: Flow cytometry ... Cells were washed with PBS and then incubated at 37 °C for 30 min in 0.5 ml of PBS containing 5 mM mβCD (Sigma) complexed with the indicated concentration of cholesterol.

    Cell Culture:

    Article Title: Uptake of Helicobacter pylori Vesicles Is Facilitated by Clathrin-Dependent and Clathrin-Independent Endocytic Pathways
    Article Snippet: Paragraph title: Cell culture, siRNA knockdown, and chemical inhibition. ... MβCD (Sigma-Aldrich) in F-12K (+FBS) was added at 4 mM or 10 mM for 15 min before vesicle addition both with and without MβCD present.

    Article Title: Epigallocatechin-3-gallate reduces inflammation induced by calcium pyrophosphate crystals in vitro
    Article Snippet: Paragraph title: THP-1 CELL CULTURE AND TREATMENT ... In some experiments, before stimulation with crystals FLS and THP-1 were pretreated for 1 h with EGCG at the indicated concentrations or for 45 min with 10 mM MβCD (Sigma-Aldrich) prepared from a 500 ng/ml PBS mother solution.

    Article Title: NOTCH3 inactivation increases triple negative breast cancer sensitivity to gefitinib by promoting EGFR tyrosine dephosphorylation and its intracellular arrest
    Article Snippet: Paragraph title: Cell culture and treatments ... Cells were treated with the following compounds: 5 mM MβCD (Sigma-Aldrich, Catalog number C4555); 3 μM GEF (Iressa, Selleckem, Houston, TX, USA; Catalog number ZD1839), 100 nM EGF Ligand (EGF; Gibco, Life Technologies, Carlsbad, CA, USA; Catalog number PHG0315); 10 μM of GSI IX (DAPT) (Calbiochem, Darmstadt, Germany; Catalog number 565770).

    Article Title: Dynamic lateral organization of opioid receptors (kappa, muwt and muN40D) in the plasma membrane at the nanoscale level, et al. Dynamic lateral organization of opioid receptors (kappa, muwt and muN40D) in the plasma membrane at the nanoscale level
    Article Snippet: Paragraph title: Cell culture, transfection, vital cell staining and immunoblotting ... Forty‐eight hours posttransfection, cells were incubated with 10 mM MβCD (Sigma) in full DMEM supplemented with 10 mM HEPES and 1 mg/mL BSA for 30 minutes at 37°C, similarly as described before.

    Article Title: Mechanistic Insights into the Cholesterol-dependent Binding of Perfringolysin O-based Probes and Cell Membranes
    Article Snippet: Flow cytometry RAW264.7 cells were cultured, non-adherent cells were eliminated, and adherent cells scraped, counted, and re-suspended in RPMI 1640 medium. .. Cells were washed with PBS and then incubated at 37 °C for 30 min in 0.5 ml of PBS containing 5 mM mβCD (Sigma) complexed with the indicated concentration of cholesterol.

    Article Title: A role for myosin-1A in the localization of a brush border disaccharidase
    Article Snippet: Paragraph title: Cell culture ... For studies involving cholesterol depletion, cells were incubated in 2% MβCD in serum-free DME (Sigma-Aldrich) for 30 min before fractionation.

    Article Title: GPI-anchored receptor clusters transiently recruit Lyn and G? for temporary cluster immobilization and Lyn activation: single-molecule tracking study 1
    Article Snippet: Paragraph title: Cell culture, drug treatments, and cDNA transfection ... Partial depletion of cholesterol in the plasma membrane was performed by incubating the cells in 4 mM MβCD (Sigma-Aldrich) at 37°C for 30 min ( ) or in 60 μg/ml saponin (Sigma-Aldrich) on ice for 15 min ( ).

    Light Microscopy:

    Article Title: A role for myosin-1A in the localization of a brush border disaccharidase
    Article Snippet: For all light microscopy studies, confluent BBEs were grown on Transwell-COL collagen-coated filters (Corning-Costar) for at least 3 wk to ensure that cells were fully differentiated. .. For studies involving cholesterol depletion, cells were incubated in 2% MβCD in serum-free DME (Sigma-Aldrich) for 30 min before fractionation.

    Generated:

    Article Title: A β-Glucan-Based Dietary Fiber Reduces Mast Cell-Induced Hyperpermeability in Ileum From Patients With Crohn’s Disease and Control Subjects
    Article Snippet: Upon M cell-transformation, monolayers were washed and preincubated with HBSS with or without the addition of 3 mM MβCD (Sigma-Aldrich) or 30 µM CPZ (Fluka) at 37°C for 20 minutes. .. The fluorescence intensity data generated from these measurements were used for subsequent statistical analysis.

    Inhibition:

    Article Title: Uptake of Helicobacter pylori Vesicles Is Facilitated by Clathrin-Dependent and Clathrin-Independent Endocytic Pathways
    Article Snippet: Paragraph title: Cell culture, siRNA knockdown, and chemical inhibition. ... MβCD (Sigma-Aldrich) in F-12K (+FBS) was added at 4 mM or 10 mM for 15 min before vesicle addition both with and without MβCD present.

    Injection:

    Article Title: Membrane stiffening by STOML3 facilitates mechanosensation in sensory neurons
    Article Snippet: .. Intraplantar injection of MβCD (Sigma, 2 mg kg−1 ) dissolved in sterile saline was performed 2 days after CCI. .. The withdrawal threshold was measured for each mouse before and at various times after injection.

    Recombinant:

    Article Title: Exosomal miR-9 released from HIV Tat stimulated astrocytes mediates microglial migration
    Article Snippet: Recombinant Tat101 was purchased from ImmunoDiagnostics (Woburn, MA). .. Chemical inhibitors, including MβCD and DMA were purchased from Sigma (St. Louis, MO, USA).

    Molecular Weight:

    Article Title: Cholesterol modulates open probability and desensitization of NMDA receptors
    Article Snippet: .. For acute cholesterol depletion by MβCD, DIV 6–7 CGCs were exposed to low-potassium medium supplemented with 5 m m MβCD (mean molecular weight 1310 g mol−1 ; Aldrich, St Louis, MO, USA) and incubated at 37°C in 5% CO2 for 1–60 min as indicated. .. Chronic cholesterol depletion by simvastatin was carried out by incubating DIV 5 CGCs in low-potassium medium supplemented with 100 n m simvastatin (Sigma) for 4 days prior to patch-clamp measurements (Zacco et al .).

    Fluorescence:

    Article Title: A β-Glucan-Based Dietary Fiber Reduces Mast Cell-Induced Hyperpermeability in Ileum From Patients With Crohn’s Disease and Control Subjects
    Article Snippet: Upon M cell-transformation, monolayers were washed and preincubated with HBSS with or without the addition of 3 mM MβCD (Sigma-Aldrich) or 30 µM CPZ (Fluka) at 37°C for 20 minutes. .. Basolateral media were collected in triplicates and β-glucan fluorescence was measured in Modulus Microplate Photometer (λex = 525nm, λem = 580–640 nm).

    Multiple Displacement Amplification:

    Article Title: NOTCH3 inactivation increases triple negative breast cancer sensitivity to gefitinib by promoting EGFR tyrosine dephosphorylation and its intracellular arrest
    Article Snippet: Cell culture and treatments Human BC MDA-MB-468, MDA-MB-231, HCC1143, and BT-549 TNBC cell lines were obtained from ATCC; HCC38, BT-20, HS578T, and MDA-MB-453 TNBC cell lines were kindly provided by Professor JV Olsen (Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Denmark). .. Cells were treated with the following compounds: 5 mM MβCD (Sigma-Aldrich, Catalog number C4555); 3 μM GEF (Iressa, Selleckem, Houston, TX, USA; Catalog number ZD1839), 100 nM EGF Ligand (EGF; Gibco, Life Technologies, Carlsbad, CA, USA; Catalog number PHG0315); 10 μM of GSI IX (DAPT) (Calbiochem, Darmstadt, Germany; Catalog number 565770).

    Article Title: Dynamic lateral organization of opioid receptors (kappa, muwt and muN40D) in the plasma membrane at the nanoscale level, et al. Dynamic lateral organization of opioid receptors (kappa, muwt and muN40D) in the plasma membrane at the nanoscale level
    Article Snippet: COS‐7, PANC‐1, MCF‐7 and MDA‐MB‐468 cells were cultured in phenol red‐free Dulbecco's modified eagle medium (DMEM) supplemented with 10% FBS, 1 mM sodium pyruvate, 100 units/mL penicillin, 100 units/mL streptomycin and 2 mM l ‐alanyl‐l ‐glutamine (full DMEM). .. Forty‐eight hours posttransfection, cells were incubated with 10 mM MβCD (Sigma) in full DMEM supplemented with 10 mM HEPES and 1 mg/mL BSA for 30 minutes at 37°C, similarly as described before.

    Isolation:

    Article Title: Cytotoxicity of purified listeriolysin O on mouse and human leukocytes and leukaemia cells
    Article Snippet: Isolation of PBMC was performed by an overlayer of peripheral blood diluted with PBS (1:1), on 5 ml of Histopaque 1077 (Sigma) preparation in a test tube and centrifugation for 30 min at 540 × g . .. For some experiments cell cultures were supplemented with the following compounds to modify the LLO activity: cholesterol (Sigma), NCS containing 150 μg/ml cholesterol (390 μM), and MβCD (Sigma).

    Transfection:

    Article Title: Dynamic lateral organization of opioid receptors (kappa, muwt and muN40D) in the plasma membrane at the nanoscale level, et al. Dynamic lateral organization of opioid receptors (kappa, muwt and muN40D) in the plasma membrane at the nanoscale level
    Article Snippet: Paragraph title: Cell culture, transfection, vital cell staining and immunoblotting ... Forty‐eight hours posttransfection, cells were incubated with 10 mM MβCD (Sigma) in full DMEM supplemented with 10 mM HEPES and 1 mg/mL BSA for 30 minutes at 37°C, similarly as described before.

    Article Title: GPI-anchored receptor clusters transiently recruit Lyn and G? for temporary cluster immobilization and Lyn activation: single-molecule tracking study 1
    Article Snippet: Paragraph title: Cell culture, drug treatments, and cDNA transfection ... Partial depletion of cholesterol in the plasma membrane was performed by incubating the cells in 4 mM MβCD (Sigma-Aldrich) at 37°C for 30 min ( ) or in 60 μg/ml saponin (Sigma-Aldrich) on ice for 15 min ( ).

    Labeling:

    Article Title: Mechanistic Insights into the Cholesterol-dependent Binding of Perfringolysin O-based Probes and Cell Membranes
    Article Snippet: Cells were washed with PBS and then incubated at 37 °C for 30 min in 0.5 ml of PBS containing 5 mM mβCD (Sigma) complexed with the indicated concentration of cholesterol. .. The cells were then washed twice with PBS and incubated at 4 °C for 20 min with 0.5 μM of the indicated PFO derivative (10% labeled with Alexa488) in 100 μl of PBS.

    Mouse Assay:

    Article Title: Membrane stiffening by STOML3 facilitates mechanosensation in sensory neurons
    Article Snippet: In brief, in deeply anaesthetized mice (isoflurane) four loose silk ligatures (4/0) were placed (with about 0.5 mm spacing) around the sciatic nerve at the level of the left mid-thigh. .. Intraplantar injection of MβCD (Sigma, 2 mg kg−1 ) dissolved in sterile saline was performed 2 days after CCI.

    Exclusion Assay:

    Article Title: NOTCH3 inactivation increases triple negative breast cancer sensitivity to gefitinib by promoting EGFR tyrosine dephosphorylation and its intracellular arrest
    Article Snippet: Cell viability was measured by the Trypan blue dye exclusion assay (Sigma-Aldrich, St Louis, MO, USA, Catalog number T8154). .. Cells were treated with the following compounds: 5 mM MβCD (Sigma-Aldrich, Catalog number C4555); 3 μM GEF (Iressa, Selleckem, Houston, TX, USA; Catalog number ZD1839), 100 nM EGF Ligand (EGF; Gibco, Life Technologies, Carlsbad, CA, USA; Catalog number PHG0315); 10 μM of GSI IX (DAPT) (Calbiochem, Darmstadt, Germany; Catalog number 565770).

    Microscopy:

    Article Title: Binding of Human Immunodeficiency Virus Type 1 to Immature Dendritic Cells Can Occur Independently of DC-SIGN and Mannose Binding C-Type Lectin Receptors via a Cholesterol-Dependent Pathway
    Article Snippet: As controls for the effective functioning of amantadine and MβCD, drug-treated DC (as described above) were incubated with fluorescein isothiocyanate (FITC)-conjugated transferrin substrate (50 μg/ml) (Sigma) for 1 h at 4°C and then shifted to 37°C for 1 h, cytospun onto coverslips, and then fixed in 4% paraformaldehyde. .. Images of the stained cells were observed and collected with the Delta Vision SA3.1 microscope (Applied Precision Inc., Issaquah, Wash.).

    Confocal Microscopy:

    Article Title: Binding of Human Immunodeficiency Virus Type 1 to Immature Dendritic Cells Can Occur Independently of DC-SIGN and Mannose Binding C-Type Lectin Receptors via a Cholesterol-Dependent Pathway
    Article Snippet: Paragraph title: Confocal microscopy. ... As controls for the effective functioning of amantadine and MβCD, drug-treated DC (as described above) were incubated with fluorescein isothiocyanate (FITC)-conjugated transferrin substrate (50 μg/ml) (Sigma) for 1 h at 4°C and then shifted to 37°C for 1 h, cytospun onto coverslips, and then fixed in 4% paraformaldehyde.

    Centrifugation:

    Article Title: Cytotoxicity of purified listeriolysin O on mouse and human leukocytes and leukaemia cells
    Article Snippet: Isolation of PBMC was performed by an overlayer of peripheral blood diluted with PBS (1:1), on 5 ml of Histopaque 1077 (Sigma) preparation in a test tube and centrifugation for 30 min at 540 × g . .. For some experiments cell cultures were supplemented with the following compounds to modify the LLO activity: cholesterol (Sigma), NCS containing 150 μg/ml cholesterol (390 μM), and MβCD (Sigma).

    Plasmid Preparation:

    Article Title: Exosomal miR-9 released from HIV Tat stimulated astrocytes mediates microglial migration
    Article Snippet: Chemical inhibitors, including MβCD and DMA were purchased from Sigma (St. Louis, MO, USA). .. The miR-9 overexpression plasmid has-miR-9-1, miR-control, and miR-9 inhibitor plasmid hsa-miR-9-5p-locker/hsa-miR-9-3p-locker as well as miR-locker-control plasmids and lentiviruses were purchased from Biosettia (San Diego, CA).

    Article Title: Binding of Human Immunodeficiency Virus Type 1 to Immature Dendritic Cells Can Occur Independently of DC-SIGN and Mannose Binding C-Type Lectin Receptors via a Cholesterol-Dependent Pathway
    Article Snippet: As controls for the effective functioning of amantadine and MβCD, drug-treated DC (as described above) were incubated with fluorescein isothiocyanate (FITC)-conjugated transferrin substrate (50 μg/ml) (Sigma) for 1 h at 4°C and then shifted to 37°C for 1 h, cytospun onto coverslips, and then fixed in 4% paraformaldehyde. .. Coverslips were rinsed extensively with PBS, stained with DAPI (4′,6′-diamidino-2-phenylindole; 0.2 μg/ml in distilled H2 O) for 5 min, washed once with distilled H2 O, dehydrated in 100% methanol for 15 min, air dried, mounted in Vectashield (Vector Laboratories, Burlingame, Calif.), and sealed with nail polish.

    Functional Assay:

    Article Title: GPI-anchored receptor clusters transiently recruit Lyn and G? for temporary cluster immobilization and Lyn activation: single-molecule tracking study 1
    Article Snippet: To examine the involvement of Lyn in STALL induction, SYF cells were transfected with the cDNA for Lyn fused with GFP at the C terminus (Lyn-GFP); this GFP construct is known to be functional ( ). .. Partial depletion of cholesterol in the plasma membrane was performed by incubating the cells in 4 mM MβCD (Sigma-Aldrich) at 37°C for 30 min ( ) or in 60 μg/ml saponin (Sigma-Aldrich) on ice for 15 min ( ).

    In Vitro:

    Article Title: Cytotoxicity of purified listeriolysin O on mouse and human leukocytes and leukaemia cells
    Article Snippet: For human peripheral blood experiments in vitro , patients gave their personal consent and the procedure was accepted by the Bioethical Commission at the Medical Center of Postgraduate Education, Warsaw (Feb 27, 2008). .. For some experiments cell cultures were supplemented with the following compounds to modify the LLO activity: cholesterol (Sigma), NCS containing 150 μg/ml cholesterol (390 μM), and MβCD (Sigma).

    Patch Clamp:

    Article Title: Cholesterol modulates open probability and desensitization of NMDA receptors
    Article Snippet: For acute cholesterol depletion by MβCD, DIV 6–7 CGCs were exposed to low-potassium medium supplemented with 5 m m MβCD (mean molecular weight 1310 g mol−1 ; Aldrich, St Louis, MO, USA) and incubated at 37°C in 5% CO2 for 1–60 min as indicated. .. Chronic cholesterol depletion by simvastatin was carried out by incubating DIV 5 CGCs in low-potassium medium supplemented with 100 n m simvastatin (Sigma) for 4 days prior to patch-clamp measurements (Zacco et al .).

    Concentration Assay:

    Article Title: Epigallocatechin-3-gallate reduces inflammation induced by calcium pyrophosphate crystals in vitro
    Article Snippet: Cells were then treated with CPP (final concentration 0.025 mg/ml; InvivoGen) for 24 h in presence or absence of EGCG (10, 50 μM) and 2% FCS. .. In some experiments, before stimulation with crystals FLS and THP-1 were pretreated for 1 h with EGCG at the indicated concentrations or for 45 min with 10 mM MβCD (Sigma-Aldrich) prepared from a 500 ng/ml PBS mother solution.

    Article Title: Mechanistic Insights into the Cholesterol-dependent Binding of Perfringolysin O-based Probes and Cell Membranes
    Article Snippet: .. Cells were washed with PBS and then incubated at 37 °C for 30 min in 0.5 ml of PBS containing 5 mM mβCD (Sigma) complexed with the indicated concentration of cholesterol. .. The cells were then washed twice with PBS and incubated at 4 °C for 20 min with 0.5 μM of the indicated PFO derivative (10% labeled with Alexa488) in 100 μl of PBS.

    Cell Counting:

    Article Title: Telocinobufagin and Marinobufagin Produce Different Effects in LLC-PK1 Cells: A Case of Functional Selectivity of Bufadienolides
    Article Snippet: Paragraph title: 4.6. Cell Counting Assay ... In 24-well plates, 1–2 × 104 LLC-PK1 cells/well were treated with 1, 10 or 100 nM CTS for 24, 48, and 72 h. In order to investigate the involvement of intracellular signaling pathways, 100 nM CTS were incubated with or without inhibitors of Src (SU6656, 10 μM; Sigma-Aldrich, St. Louis, MO, USA), ERK1/2-MEK1/2 (U0126, 10 μM; Cell Signaling Technology, Danvers, MA, USA), PI3K ( , 5 μM; Cell Signaling Technology, Danvers, MA, USA), p38 (SB202190, 10 μM; Cell Signaling Technology, Danvers, MA, USA), JNK1/2 (SP600125, 1.5 μM; Cell Signaling Technology, Danvers, MA, USA), or GSK-3β (BIO, 0.5, 1, and 5 μM; Sigma-Aldrich, St. Louis, MO, USA) for 72 h. For MβCD (Sigma-Aldrich, St. Louis, MO, USA) assay, cells were pretreated with 10 mM MβCD for 30 min and then 1 mM MβCD + 100 nM TCB in 2.5% FBS for 72 h. At these time points, two wells from each group were trypsinized and the number of Trypan blue-viable cells was counted in Neubauer chamber (hemocytometer).

    Fractionation:

    Article Title: A role for myosin-1A in the localization of a brush border disaccharidase
    Article Snippet: .. For studies involving cholesterol depletion, cells were incubated in 2% MβCD in serum-free DME (Sigma-Aldrich) for 30 min before fractionation. .. Cloning of GFP-M1A-tail and SINT-YFP fusion constructs The GFP-M1A-tail and SINT-YFP fusion constructs were assembled using standard molecular biological techniques.

    Staining:

    Article Title: Dynamic lateral organization of opioid receptors (kappa, muwt and muN40D) in the plasma membrane at the nanoscale level, et al. Dynamic lateral organization of opioid receptors (kappa, muwt and muN40D) in the plasma membrane at the nanoscale level
    Article Snippet: Paragraph title: Cell culture, transfection, vital cell staining and immunoblotting ... Forty‐eight hours posttransfection, cells were incubated with 10 mM MβCD (Sigma) in full DMEM supplemented with 10 mM HEPES and 1 mg/mL BSA for 30 minutes at 37°C, similarly as described before.

    Article Title: Binding of Human Immunodeficiency Virus Type 1 to Immature Dendritic Cells Can Occur Independently of DC-SIGN and Mannose Binding C-Type Lectin Receptors via a Cholesterol-Dependent Pathway
    Article Snippet: As controls for the effective functioning of amantadine and MβCD, drug-treated DC (as described above) were incubated with fluorescein isothiocyanate (FITC)-conjugated transferrin substrate (50 μg/ml) (Sigma) for 1 h at 4°C and then shifted to 37°C for 1 h, cytospun onto coverslips, and then fixed in 4% paraformaldehyde. .. Coverslips were rinsed extensively with PBS, stained with DAPI (4′,6′-diamidino-2-phenylindole; 0.2 μg/ml in distilled H2 O) for 5 min, washed once with distilled H2 O, dehydrated in 100% methanol for 15 min, air dried, mounted in Vectashield (Vector Laboratories, Burlingame, Calif.), and sealed with nail polish.

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  • 95
    Millipore cholesterol mβcd
    Adding cholesterol reverses antiinflammatory effect of AIBP. MH-S cells were preincubated for 2 hours with 50 μg/ml surfactant in the presence or absence of 0.2 μg/ml AIBP, followed by a 1-hour treatment with or without 50 μg/ml cholesterol <t>methyl-β-cyclodextrin</t> (chol-βCD). The cells were then stimulated with 10 ng/ml LPS for 30 minutes. ( A ) Representative immunoblot. ( B ) Quantitative data for phospho-p65, phospho-ERK1/2, and phospho-p38. Mean ± SEM; n = 5; * P
    Cholesterol Mβcd, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cholesterol mβcd/product/Millipore
    Average 95 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    cholesterol mβcd - by Bioz Stars, 2020-02
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    77
    Millipore meβcd
    Cortical 45-kDa Gαs is contained in low-density membrane and is reduced after treatment of oocytes with progesterone or <t>MeβCD</t>
    Meβcd, supplied by Millipore, used in various techniques. Bioz Stars score: 77/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/meβcd/product/Millipore
    Average 77 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    meβcd - by Bioz Stars, 2020-02
    77/100 stars
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    79
    Millipore mβcd powder
    Recovery of dysregulated protein signature in NPC1 fibroblasts upon <t>MβCD</t> treatment. (A) Heat map showing the identified and quantified proteins that are differentially expressed in NPC1 fibroblast cells, comparing with WT, and reversed by MβCD-1, MβCD-2, or MβCD-3 treatments. The color key indicates the relative abundance of proteins (0–1.0) across five samples. Relative protein levels of NPC2 (B) , USE1 (C) , VAMP7 (D) , GABARAP (E) , NSDHL (F) , and DHCR24 (G) . CD, cyclodextrin; ppm, part per million.
    Mβcd Powder, supplied by Millipore, used in various techniques. Bioz Stars score: 79/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mβcd powder/product/Millipore
    Average 79 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mβcd powder - by Bioz Stars, 2020-02
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      Buy from Supplier

    Image Search Results


    Adding cholesterol reverses antiinflammatory effect of AIBP. MH-S cells were preincubated for 2 hours with 50 μg/ml surfactant in the presence or absence of 0.2 μg/ml AIBP, followed by a 1-hour treatment with or without 50 μg/ml cholesterol methyl-β-cyclodextrin (chol-βCD). The cells were then stimulated with 10 ng/ml LPS for 30 minutes. ( A ) Representative immunoblot. ( B ) Quantitative data for phospho-p65, phospho-ERK1/2, and phospho-p38. Mean ± SEM; n = 5; * P

    Journal: JCI Insight

    Article Title: AIBP augments cholesterol efflux from alveolar macrophages to surfactant and reduces acute lung inflammation

    doi: 10.1172/jci.insight.120519

    Figure Lengend Snippet: Adding cholesterol reverses antiinflammatory effect of AIBP. MH-S cells were preincubated for 2 hours with 50 μg/ml surfactant in the presence or absence of 0.2 μg/ml AIBP, followed by a 1-hour treatment with or without 50 μg/ml cholesterol methyl-β-cyclodextrin (chol-βCD). The cells were then stimulated with 10 ng/ml LPS for 30 minutes. ( A ) Representative immunoblot. ( B ) Quantitative data for phospho-p65, phospho-ERK1/2, and phospho-p38. Mean ± SEM; n = 5; * P

    Article Snippet: In some experiments, cells were incubated for 1 hour with or without 50 μg/ml cholesterol-MβCD (MilliporeSigma, C4951).

    Techniques:

    Macrophages release particles enriched in cholesterol. ( A ) Macrophages were loaded with cholesterol/MβCD. SEM and nanoSIMS images of the macrophage after a short incubation with [ 15 N]ALO-D4. The SEM image shows a lawn of particles outside the macrophage; the nanoSIMS image (scaled at two different settings) reveals binding of [ 15 N]ALO-D4 to the particles, indicating that they contain accessible cholesterol. The boxed regions are shown below at higher magnification, again showing binding of [ 15 N]ALO-D4 to macrophage-derived particles. (Scale bars, 5 μm.) The bar graph shows 15 N/ 14 N levels for the cell body and particles of two cells (60 particles were quantified). The y axis starts at 0.0037, the natural abundance of 15 N. Data are shown as mean ± SD. ( B ) Macrophages were loaded with acetyl-LDL. Particles released by macrophages that had been loaded with acetyl-LDL (50 μg/mL) (yellow arrows) are visible in secondary electron (SE) and 12 C 14 N − nanoSIMS images. Composite 12 C 14 N − or secondary electron (SE) (gray) and 15 N/ 14 N ratio (red) images show binding of [ 15 N]ALO-D4 to particles. (Scale bars, 2 μm.)

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Macrophages release plasma membrane-derived particles rich in accessible cholesterol

    doi: 10.1073/pnas.1810724115

    Figure Lengend Snippet: Macrophages release particles enriched in cholesterol. ( A ) Macrophages were loaded with cholesterol/MβCD. SEM and nanoSIMS images of the macrophage after a short incubation with [ 15 N]ALO-D4. The SEM image shows a lawn of particles outside the macrophage; the nanoSIMS image (scaled at two different settings) reveals binding of [ 15 N]ALO-D4 to the particles, indicating that they contain accessible cholesterol. The boxed regions are shown below at higher magnification, again showing binding of [ 15 N]ALO-D4 to macrophage-derived particles. (Scale bars, 5 μm.) The bar graph shows 15 N/ 14 N levels for the cell body and particles of two cells (60 particles were quantified). The y axis starts at 0.0037, the natural abundance of 15 N. Data are shown as mean ± SD. ( B ) Macrophages were loaded with acetyl-LDL. Particles released by macrophages that had been loaded with acetyl-LDL (50 μg/mL) (yellow arrows) are visible in secondary electron (SE) and 12 C 14 N − nanoSIMS images. Composite 12 C 14 N − or secondary electron (SE) (gray) and 15 N/ 14 N ratio (red) images show binding of [ 15 N]ALO-D4 to particles. (Scale bars, 2 μm.)

    Article Snippet: Macrophages were loaded with 20 μL/mL of [13 C]cholesterol/MβCD (final cholesterol concentration, 50 μM) in macrophage medium containing 0.1% LPDS, 50 μM mevastatin (Calbiochem), and 50 μM mevalonolactone (Sigma) for 24 h at 37 °C.

    Techniques: Incubation, Binding Assay, Derivative Assay

    Cortical 45-kDa Gαs is contained in low-density membrane and is reduced after treatment of oocytes with progesterone or MeβCD

    Journal: Developmental biology

    Article Title: Activation of the progesterone signaling pathway by methyl-?-cyclodextrin or steroid in Xenopus laevis oocytes involves release of 45-kDa G?s

    doi: 10.1016/j.ydbio.2008.07.031

    Figure Lengend Snippet: Cortical 45-kDa Gαs is contained in low-density membrane and is reduced after treatment of oocytes with progesterone or MeβCD

    Article Snippet: Treatment of oocytes with progesterone or MeβCD reduced the apparent amounts of 45-kDa Gαs detected in oocyte cortices with the Calbiochem anti-Gαs antibody ( ).

    Techniques:

    Progesterone- and MeβCD-stimulated increases in oocyte 39-kDa Mos are inhibited by cycloheximide

    Journal: Developmental biology

    Article Title: Activation of the progesterone signaling pathway by methyl-?-cyclodextrin or steroid in Xenopus laevis oocytes involves release of 45-kDa G?s

    doi: 10.1016/j.ydbio.2008.07.031

    Figure Lengend Snippet: Progesterone- and MeβCD-stimulated increases in oocyte 39-kDa Mos are inhibited by cycloheximide

    Article Snippet: Treatment of oocytes with progesterone or MeβCD reduced the apparent amounts of 45-kDa Gαs detected in oocyte cortices with the Calbiochem anti-Gαs antibody ( ).

    Techniques:

    The steroid synthesis inhibitor, aminoglutethimide, does not block oocyte maturation stimulated MeβCD

    Journal: Developmental biology

    Article Title: Activation of the progesterone signaling pathway by methyl-?-cyclodextrin or steroid in Xenopus laevis oocytes involves release of 45-kDa G?s

    doi: 10.1016/j.ydbio.2008.07.031

    Figure Lengend Snippet: The steroid synthesis inhibitor, aminoglutethimide, does not block oocyte maturation stimulated MeβCD

    Article Snippet: Treatment of oocytes with progesterone or MeβCD reduced the apparent amounts of 45-kDa Gαs detected in oocyte cortices with the Calbiochem anti-Gαs antibody ( ).

    Techniques: Blocking Assay

    Follicle cells are not required for phosphorylation of oocyte MAPK or GVBD induced by MeβCD

    Journal: Developmental biology

    Article Title: Activation of the progesterone signaling pathway by methyl-?-cyclodextrin or steroid in Xenopus laevis oocytes involves release of 45-kDa G?s

    doi: 10.1016/j.ydbio.2008.07.031

    Figure Lengend Snippet: Follicle cells are not required for phosphorylation of oocyte MAPK or GVBD induced by MeβCD

    Article Snippet: Treatment of oocytes with progesterone or MeβCD reduced the apparent amounts of 45-kDa Gαs detected in oocyte cortices with the Calbiochem anti-Gαs antibody ( ).

    Techniques:

    Cycloheximide blocks MAPK phosphorylation and GVBD in response to MeβCD

    Journal: Developmental biology

    Article Title: Activation of the progesterone signaling pathway by methyl-?-cyclodextrin or steroid in Xenopus laevis oocytes involves release of 45-kDa G?s

    doi: 10.1016/j.ydbio.2008.07.031

    Figure Lengend Snippet: Cycloheximide blocks MAPK phosphorylation and GVBD in response to MeβCD

    Article Snippet: Treatment of oocytes with progesterone or MeβCD reduced the apparent amounts of 45-kDa Gαs detected in oocyte cortices with the Calbiochem anti-Gαs antibody ( ).

    Techniques:

    Recovery of dysregulated protein signature in NPC1 fibroblasts upon MβCD treatment. (A) Heat map showing the identified and quantified proteins that are differentially expressed in NPC1 fibroblast cells, comparing with WT, and reversed by MβCD-1, MβCD-2, or MβCD-3 treatments. The color key indicates the relative abundance of proteins (0–1.0) across five samples. Relative protein levels of NPC2 (B) , USE1 (C) , VAMP7 (D) , GABARAP (E) , NSDHL (F) , and DHCR24 (G) . CD, cyclodextrin; ppm, part per million.

    Journal: Assay and Drug Development Technologies

    Article Title: Analytical Characterization of Methyl-β-Cyclodextrin for Pharmacological Activity to Reduce Lysosomal Cholesterol Accumulation in Niemann-Pick Disease Type C1 Cells

    doi: 10.1089/adt.2017.774

    Figure Lengend Snippet: Recovery of dysregulated protein signature in NPC1 fibroblasts upon MβCD treatment. (A) Heat map showing the identified and quantified proteins that are differentially expressed in NPC1 fibroblast cells, comparing with WT, and reversed by MβCD-1, MβCD-2, or MβCD-3 treatments. The color key indicates the relative abundance of proteins (0–1.0) across five samples. Relative protein levels of NPC2 (B) , USE1 (C) , VAMP7 (D) , GABARAP (E) , NSDHL (F) , and DHCR24 (G) . CD, cyclodextrin; ppm, part per million.

    Article Snippet: Initially, 0.4 mg of MβCD powder was dissolved in 3 mL of deionized Millipore (Sigma-Aldrich, St. Louis, MO) water as the stock solution (100 μM).

    Techniques:

    Effects of different sources of MβCDs on reducing lysosome size in NPC1 fibroblasts. NPC1 patient skin fibroblasts (GM03123) and WT control (GM05659) were untreated or treated with MβCD (0.4–300 μM) for 4 days, after which LysoTracker ® staining was performed. (A) Images of LysoTracker staining on NPC1 fibroblasts. Treatment with 300 and 11 μM of MβCD-3 significantly reduced the cholesterol accumulation in NPC1 patient skin fibroblasts, while there were no significant effects observed from the other two batches of MβCDs (MβCD-1 and MβCD-2). LysoTracker red stains cellular acidic compartments to visualize enlarged lysosomes, and Hoechst ( blue ) stains nuclei. (B) Treatment with MβCD-3 (300 μM) significantly reduced the lysosome size in the NPC1 patient fibroblast compared with the other two batches of MβCDs (MβCD-1 and MβCD-2). (C) Dose–response curve of different sources of MβCDs on NPC1 patient fibroblasts. MβCD-3 showed concentration-dependent impact on lysosome size of NPC1 fibroblasts, while there were no significant effects observed from the other two batches of MβCDs.

    Journal: Assay and Drug Development Technologies

    Article Title: Analytical Characterization of Methyl-β-Cyclodextrin for Pharmacological Activity to Reduce Lysosomal Cholesterol Accumulation in Niemann-Pick Disease Type C1 Cells

    doi: 10.1089/adt.2017.774

    Figure Lengend Snippet: Effects of different sources of MβCDs on reducing lysosome size in NPC1 fibroblasts. NPC1 patient skin fibroblasts (GM03123) and WT control (GM05659) were untreated or treated with MβCD (0.4–300 μM) for 4 days, after which LysoTracker ® staining was performed. (A) Images of LysoTracker staining on NPC1 fibroblasts. Treatment with 300 and 11 μM of MβCD-3 significantly reduced the cholesterol accumulation in NPC1 patient skin fibroblasts, while there were no significant effects observed from the other two batches of MβCDs (MβCD-1 and MβCD-2). LysoTracker red stains cellular acidic compartments to visualize enlarged lysosomes, and Hoechst ( blue ) stains nuclei. (B) Treatment with MβCD-3 (300 μM) significantly reduced the lysosome size in the NPC1 patient fibroblast compared with the other two batches of MβCDs (MβCD-1 and MβCD-2). (C) Dose–response curve of different sources of MβCDs on NPC1 patient fibroblasts. MβCD-3 showed concentration-dependent impact on lysosome size of NPC1 fibroblasts, while there were no significant effects observed from the other two batches of MβCDs.

    Article Snippet: Initially, 0.4 mg of MβCD powder was dissolved in 3 mL of deionized Millipore (Sigma-Aldrich, St. Louis, MO) water as the stock solution (100 μM).

    Techniques: Staining, Concentration Assay

    Mass spectrometry on MβCD. Mass spectra of MβCD-1, -2, and -3 show cluster signals of sodium adduct ions of 6–12 different mixtures of methyl-substituted β-cyclodextrin molecules. Methylation number can be easily determined by an inclement of mass unit (m/z) 14 as shown. The mass spectrometry peak heights are proportional to molecular distributions (or amounts) of various methyl-substituted β-cyclodextrins (summarized in Table 1 ). Abundance of lower number of methyl substitution of MβCD is shown in (A) (MβCD-1), while middle substitution is in (B) (MβCD-2) and higher substitution is in (C) (MβCD-3). 10-Me, 10 methylation to β-cyclodextrin molecule.

    Journal: Assay and Drug Development Technologies

    Article Title: Analytical Characterization of Methyl-β-Cyclodextrin for Pharmacological Activity to Reduce Lysosomal Cholesterol Accumulation in Niemann-Pick Disease Type C1 Cells

    doi: 10.1089/adt.2017.774

    Figure Lengend Snippet: Mass spectrometry on MβCD. Mass spectra of MβCD-1, -2, and -3 show cluster signals of sodium adduct ions of 6–12 different mixtures of methyl-substituted β-cyclodextrin molecules. Methylation number can be easily determined by an inclement of mass unit (m/z) 14 as shown. The mass spectrometry peak heights are proportional to molecular distributions (or amounts) of various methyl-substituted β-cyclodextrins (summarized in Table 1 ). Abundance of lower number of methyl substitution of MβCD is shown in (A) (MβCD-1), while middle substitution is in (B) (MβCD-2) and higher substitution is in (C) (MβCD-3). 10-Me, 10 methylation to β-cyclodextrin molecule.

    Article Snippet: Initially, 0.4 mg of MβCD powder was dissolved in 3 mL of deionized Millipore (Sigma-Aldrich, St. Louis, MO) water as the stock solution (100 μM).

    Techniques: Mass Spectrometry, Methylation

    Chemical structure of β-cyclodextrin. (A) Chemical representation of methyl-β-cyclodextrin (MβCD), which comprises seven glucopyranose units. (B) Three-dimensional representation of the toroid structure of cyclodextrin consisting of a hydrophilic exterior and hydrophobic interior.

    Journal: Assay and Drug Development Technologies

    Article Title: Analytical Characterization of Methyl-β-Cyclodextrin for Pharmacological Activity to Reduce Lysosomal Cholesterol Accumulation in Niemann-Pick Disease Type C1 Cells

    doi: 10.1089/adt.2017.774

    Figure Lengend Snippet: Chemical structure of β-cyclodextrin. (A) Chemical representation of methyl-β-cyclodextrin (MβCD), which comprises seven glucopyranose units. (B) Three-dimensional representation of the toroid structure of cyclodextrin consisting of a hydrophilic exterior and hydrophobic interior.

    Article Snippet: Initially, 0.4 mg of MβCD powder was dissolved in 3 mL of deionized Millipore (Sigma-Aldrich, St. Louis, MO) water as the stock solution (100 μM).

    Techniques:

    Effects of different sources of MβCDs on reducing cholesterol accumulation in NPC1 fibroblasts. NPC1 patient skin fibroblasts (GM03123) and WT control (GM05659) were untreated or treated with MβCD (0.4–300 μM) for 4 days; filipin staining was then performed. (A) Images of filipin staining on NPC1 fibroblasts. Treatment with 300, 11 μM of MβCD-3 significantly reduced cholesterol accumulation in NPC1 patient skin fibroblasts, while the other two batches of MβCDs (MβCD-1 and MβCD-2) showed much weaker effects on cholesterol accumulation in NPC1 patient fibroblasts. Filipin ( green ) stains the intracellular cholesterol-laden domains, and EthD-1 ( red ) stains nuclei. (B) Treatment with MβCD-3 (300 μM) significantly reduced cholesterol accumulation in the NPC1 patient fibroblast compared with the other two batches of MβCDs (MβCD-1 and MβCD-2). (C) Dose–response curve of different sources of MβCDs on cholesterol accumulation in NPC1 patient fibroblasts. EthD-1, ethidium homodimer; NPC1, Niemann-Pick disease type C1; WT, wild-type.

    Journal: Assay and Drug Development Technologies

    Article Title: Analytical Characterization of Methyl-β-Cyclodextrin for Pharmacological Activity to Reduce Lysosomal Cholesterol Accumulation in Niemann-Pick Disease Type C1 Cells

    doi: 10.1089/adt.2017.774

    Figure Lengend Snippet: Effects of different sources of MβCDs on reducing cholesterol accumulation in NPC1 fibroblasts. NPC1 patient skin fibroblasts (GM03123) and WT control (GM05659) were untreated or treated with MβCD (0.4–300 μM) for 4 days; filipin staining was then performed. (A) Images of filipin staining on NPC1 fibroblasts. Treatment with 300, 11 μM of MβCD-3 significantly reduced cholesterol accumulation in NPC1 patient skin fibroblasts, while the other two batches of MβCDs (MβCD-1 and MβCD-2) showed much weaker effects on cholesterol accumulation in NPC1 patient fibroblasts. Filipin ( green ) stains the intracellular cholesterol-laden domains, and EthD-1 ( red ) stains nuclei. (B) Treatment with MβCD-3 (300 μM) significantly reduced cholesterol accumulation in the NPC1 patient fibroblast compared with the other two batches of MβCDs (MβCD-1 and MβCD-2). (C) Dose–response curve of different sources of MβCDs on cholesterol accumulation in NPC1 patient fibroblasts. EthD-1, ethidium homodimer; NPC1, Niemann-Pick disease type C1; WT, wild-type.

    Article Snippet: Initially, 0.4 mg of MβCD powder was dissolved in 3 mL of deionized Millipore (Sigma-Aldrich, St. Louis, MO) water as the stock solution (100 μM).

    Techniques: Staining, Ethidium Homodimer Assay

    Effect of different sources of MβCDs on cholesterol accumulation in NPC1 fibroblasts. NPC1 patient skin fibroblasts (GM03123) were treated with MβCD (0.4–300 μM) for 4 days, followed by an Amplex ® Red cholesterol assay. (A) Dose–response curve of different sources of MβCDs on NPC1 patient fibroblasts on cholesterol accumulation in NPC1 fibroblasts. MβCD-3 showed concentration-dependent manner on cholesterol accumulation in NPC1 fibroblasts GM03123, while there were much weaker effects observed with the other two batches of MβCDs. The maximum inhibitory effect of MβCD-3 is about 46.0% compared with MβCD-1, 15.2%, and MβCD-2, 19.5%. (B) Treatment with 300 μM of MβCD-3 significantly reduced cholesterol accumulation in NPC1 fibroblasts compared with the other two sources of MβCDs. (C) Cytotoxicity (ATPlite assay) of different sources of MβCDs on NPC1 fibroblasts. NPC1 patient skin fibroblasts (GM03123) were untreated or treated with MβCD (0.4–300 μM) for 4 days and ATPlite assay was performed to evaluate the cytotoxic effects of different sources of MβCDs on the fibroblasts. There were no significant cytotoxic effects observed within the test range of 0.4–300 μM of MβCD and the cell viability level was generally above 90% after 4 days of treatment.

    Journal: Assay and Drug Development Technologies

    Article Title: Analytical Characterization of Methyl-β-Cyclodextrin for Pharmacological Activity to Reduce Lysosomal Cholesterol Accumulation in Niemann-Pick Disease Type C1 Cells

    doi: 10.1089/adt.2017.774

    Figure Lengend Snippet: Effect of different sources of MβCDs on cholesterol accumulation in NPC1 fibroblasts. NPC1 patient skin fibroblasts (GM03123) were treated with MβCD (0.4–300 μM) for 4 days, followed by an Amplex ® Red cholesterol assay. (A) Dose–response curve of different sources of MβCDs on NPC1 patient fibroblasts on cholesterol accumulation in NPC1 fibroblasts. MβCD-3 showed concentration-dependent manner on cholesterol accumulation in NPC1 fibroblasts GM03123, while there were much weaker effects observed with the other two batches of MβCDs. The maximum inhibitory effect of MβCD-3 is about 46.0% compared with MβCD-1, 15.2%, and MβCD-2, 19.5%. (B) Treatment with 300 μM of MβCD-3 significantly reduced cholesterol accumulation in NPC1 fibroblasts compared with the other two sources of MβCDs. (C) Cytotoxicity (ATPlite assay) of different sources of MβCDs on NPC1 fibroblasts. NPC1 patient skin fibroblasts (GM03123) were untreated or treated with MβCD (0.4–300 μM) for 4 days and ATPlite assay was performed to evaluate the cytotoxic effects of different sources of MβCDs on the fibroblasts. There were no significant cytotoxic effects observed within the test range of 0.4–300 μM of MβCD and the cell viability level was generally above 90% after 4 days of treatment.

    Article Snippet: Initially, 0.4 mg of MβCD powder was dissolved in 3 mL of deionized Millipore (Sigma-Aldrich, St. Louis, MO) water as the stock solution (100 μM).

    Techniques: Amplex Red Cholesterol Assay, Concentration Assay