Structured Review

Merck & Co filipin iii
Filipin Iii, supplied by Merck & Co, used in various techniques. Bioz Stars score: 80/100, based on 0 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/filipin iii/product/Merck & Co
Average 80 stars, based on 1 article reviews
Price from $9.99 to $1999.99
filipin iii - by Bioz Stars, 2020-04
80/100 stars

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Concentration Assay:

Article Title: Cholesterol restricts lymphotoxin β receptor-triggered NF-κB signaling
Article Snippet: .. Filipin III (F4767, Merck) was used at a concentration of 1 μg/ml medium without FBS. .. Cholesterol (C3045, Merck) was dissolved in ethanol and then complexed with MβCD to concentration (50 mM MβCD: 5 mM Cholesterol).

Article Title: Cholesterol restricts lymphotoxin β receptor-triggered NF-κB signaling
Article Snippet: .. Cholesterol sequestration was performed using filipin III (F4767, Merck) at 1 μg/ml concentration in medium without serum. ..

Article Title: Cholesterol restricts lymphotoxin β receptor-triggered NF-κB signaling
Article Snippet: .. Following stimulation with LTβR agonist was carried out in the absence of filipin for next 0.5 h and 1 h. Acute cholesterol depletion was performed using MβCD (C4555, Merck) at a concentration of 5 mM (short treatments – 0.5, 1 or 4 h) or 2.5 mM (long treatment – 6, or 8 h). .. Chronic cholesterol depletion was achieved by incubating the cells in delipidated medium containing 30 μM simvastatin (S6196, Merck) for 48 h. Then LTβR stimulation was carried out in the same medium supplemented with LTβR agonist or lymphotoxin α1β2.

Quantitative RT-PCR:

Article Title: Cholesterol restricts lymphotoxin β receptor-triggered NF-κB signaling
Article Snippet: Cholesterol sequestration/depletion experiments Two days before experiment cells were seeded on 24-, 12-, 6-well plate or 10 cm dishes (4.5 × 104 , 10 × 104 , 25 × 104 , 200 × 104 , respectively) depending on a type of assay (microscopy, Western blotting/ qRT-PCR, immunoprecipitation). .. Cholesterol sequestration was performed using filipin III (F4767, Merck) at 1 μg/ml concentration in medium without serum.

Immunoprecipitation:

Article Title: Cholesterol restricts lymphotoxin β receptor-triggered NF-κB signaling
Article Snippet: For immunoprecipitation, goat agonistic anti-LTβR (AF629, R & D Systems) and control goat IgG (I5256, Sigma-Aldrich) were used. .. Filipin III (F4767, Merck) was used at a concentration of 1 μg/ml medium without FBS.

Article Title: Cholesterol restricts lymphotoxin β receptor-triggered NF-κB signaling
Article Snippet: Cholesterol sequestration/depletion experiments Two days before experiment cells were seeded on 24-, 12-, 6-well plate or 10 cm dishes (4.5 × 104 , 10 × 104 , 25 × 104 , 200 × 104 , respectively) depending on a type of assay (microscopy, Western blotting/ qRT-PCR, immunoprecipitation). .. Cholesterol sequestration was performed using filipin III (F4767, Merck) at 1 μg/ml concentration in medium without serum.

Microscopy:

Article Title: Cholesterol restricts lymphotoxin β receptor-triggered NF-κB signaling
Article Snippet: Cholesterol sequestration/depletion experiments Two days before experiment cells were seeded on 24-, 12-, 6-well plate or 10 cm dishes (4.5 × 104 , 10 × 104 , 25 × 104 , 200 × 104 , respectively) depending on a type of assay (microscopy, Western blotting/ qRT-PCR, immunoprecipitation). .. Cholesterol sequestration was performed using filipin III (F4767, Merck) at 1 μg/ml concentration in medium without serum.

Western Blot:

Article Title: Cholesterol restricts lymphotoxin β receptor-triggered NF-κB signaling
Article Snippet: Cholesterol sequestration/depletion experiments Two days before experiment cells were seeded on 24-, 12-, 6-well plate or 10 cm dishes (4.5 × 104 , 10 × 104 , 25 × 104 , 200 × 104 , respectively) depending on a type of assay (microscopy, Western blotting/ qRT-PCR, immunoprecipitation). .. Cholesterol sequestration was performed using filipin III (F4767, Merck) at 1 μg/ml concentration in medium without serum.

Recombinant:

Article Title: Cholesterol restricts lymphotoxin β receptor-triggered NF-κB signaling
Article Snippet: Anti-LTβR agonistic antibody (AF629, R & D Systems) and human recombinant lymphotoxin α1β2 (8884-LY-025/CF, R & D Systems) were used at concentration 0.2 μg/ml. .. Filipin III (F4767, Merck) was used at a concentration of 1 μg/ml medium without FBS.

Immunofluorescence:

Article Title: Cholesterol restricts lymphotoxin β receptor-triggered NF-κB signaling
Article Snippet: Secondary antibodies used for immunofluorescence: Alexa Fluor 488-, 555-, 647-conjugated anti-goat, anti-mouse and anti-rabbit (Thermo Fisher Scientific) were diluted 1:500. .. Filipin III (F4767, Merck) was used at a concentration of 1 μg/ml medium without FBS.

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    Merck & Co filipin
    Sequestration of the plasma membrane pool of cholesterol by <t>filipin</t> enhances LTβR-triggered activity of the NF-κB pathway and reduces internalization of the ligand-bound receptor. a Lysates of A549 cells preincubated for 0.5 h with filipin or vehicle and stimulated for 0.5 or 1 h with LTβR agonist (Ago) were analyzed by Western blotting with antibodies against the indicated proteins. Vinculin was used as a loading control. Graphs show densitometric analysis for the indicated proteins from Western blotting (protein levels normalized to vinculin). Values are presented as fold change versus controls - unstimulated and untreated cells (black bars). Data represent the means ± SEM, n ≥ 5; ns - P > 0.05; * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001 by one sample t-test (in grey), Student’s t - test (in black). b Immunofluorescence staining of ligand-bound LTβR and EEA1 upon 0.5 h stimulation with LTβR agonist in A549 cells preincubated for 0.5 h with either vehicle (Veh.) or filipin. Insets: magnified views of boxed regions in the main images. Scale bars, 20 μm. c Analysis of integral intensity and number of LTβR- and EEA1-positive vesicles in cells treated as in B. Values are presented as fold change versus control - vehicle-treated cells marked as a black line, set as 1. Data represent the means ± SEM, n = 5. ns - P > 0.05; * P ≤ 0.05; ***P ≤ 0.001 by one sample t-test
    Filipin, supplied by Merck & Co, used in various techniques. Bioz Stars score: 80/100, based on 0 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/filipin/product/Merck & Co
    Average 80 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    filipin - by Bioz Stars, 2020-04
    80/100 stars
      Buy from Supplier

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    Sequestration of the plasma membrane pool of cholesterol by filipin enhances LTβR-triggered activity of the NF-κB pathway and reduces internalization of the ligand-bound receptor. a Lysates of A549 cells preincubated for 0.5 h with filipin or vehicle and stimulated for 0.5 or 1 h with LTβR agonist (Ago) were analyzed by Western blotting with antibodies against the indicated proteins. Vinculin was used as a loading control. Graphs show densitometric analysis for the indicated proteins from Western blotting (protein levels normalized to vinculin). Values are presented as fold change versus controls - unstimulated and untreated cells (black bars). Data represent the means ± SEM, n ≥ 5; ns - P > 0.05; * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001 by one sample t-test (in grey), Student’s t - test (in black). b Immunofluorescence staining of ligand-bound LTβR and EEA1 upon 0.5 h stimulation with LTβR agonist in A549 cells preincubated for 0.5 h with either vehicle (Veh.) or filipin. Insets: magnified views of boxed regions in the main images. Scale bars, 20 μm. c Analysis of integral intensity and number of LTβR- and EEA1-positive vesicles in cells treated as in B. Values are presented as fold change versus control - vehicle-treated cells marked as a black line, set as 1. Data represent the means ± SEM, n = 5. ns - P > 0.05; * P ≤ 0.05; ***P ≤ 0.001 by one sample t-test

    Journal: Cell Communication and Signaling : CCS

    Article Title: Cholesterol restricts lymphotoxin β receptor-triggered NF-κB signaling

    doi: 10.1186/s12964-019-0460-1

    Figure Lengend Snippet: Sequestration of the plasma membrane pool of cholesterol by filipin enhances LTβR-triggered activity of the NF-κB pathway and reduces internalization of the ligand-bound receptor. a Lysates of A549 cells preincubated for 0.5 h with filipin or vehicle and stimulated for 0.5 or 1 h with LTβR agonist (Ago) were analyzed by Western blotting with antibodies against the indicated proteins. Vinculin was used as a loading control. Graphs show densitometric analysis for the indicated proteins from Western blotting (protein levels normalized to vinculin). Values are presented as fold change versus controls - unstimulated and untreated cells (black bars). Data represent the means ± SEM, n ≥ 5; ns - P > 0.05; * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001 by one sample t-test (in grey), Student’s t - test (in black). b Immunofluorescence staining of ligand-bound LTβR and EEA1 upon 0.5 h stimulation with LTβR agonist in A549 cells preincubated for 0.5 h with either vehicle (Veh.) or filipin. Insets: magnified views of boxed regions in the main images. Scale bars, 20 μm. c Analysis of integral intensity and number of LTβR- and EEA1-positive vesicles in cells treated as in B. Values are presented as fold change versus control - vehicle-treated cells marked as a black line, set as 1. Data represent the means ± SEM, n = 5. ns - P > 0.05; * P ≤ 0.05; ***P ≤ 0.001 by one sample t-test

    Article Snippet: Following stimulation with LTβR agonist was carried out in the absence of filipin for next 0.5 h and 1 h. Acute cholesterol depletion was performed using MβCD (C4555, Merck) at a concentration of 5 mM (short treatments – 0.5, 1 or 4 h) or 2.5 mM (long treatment – 6, or 8 h).

    Techniques: Activity Assay, Western Blot, Immunofluorescence, Staining