egta ethylene glycol bis 2 aminoethylether n  (Millipore)

 
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    Name:
    Ethylene glycol bis 2 aminoethylether N N N N tetraacetic acid
    Description:

    Catalog Number:
    e3889
    Price:
    None
    Applications:
    A chelating agent useful for the determination of calcium in the presence of magnesium.
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    Structured Review

    Millipore egta ethylene glycol bis 2 aminoethylether n
    Ethylene glycol bis 2 aminoethylether N N N N tetraacetic acid

    https://www.bioz.com/result/egta ethylene glycol bis 2 aminoethylether n/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    egta ethylene glycol bis 2 aminoethylether n - by Bioz Stars, 2021-03
    99/100 stars

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    Related Articles

    Blocking Assay:

    Article Title: Spexin Enhances Bowel Movement through Activating L-type Voltage-dependent Calcium Channel via Galanin Receptor 2 in Mice
    Article Snippet: Acetylcholine chloride (ACh)(Cat. no. A2661; Sigma, St. Louis, MO USA) and KCl (Cat. no. P9333; Sigma-Aldrich, St. Louis, MO USA) were used as positive controls. .. To examine the possible mechanisms of spexin on bowel movement, TTX (cat. no. 1069; Tocris Bioscience, Bristol, UK), EGTA (cat. no. E3889; Sigma-Aldrich, St. Louis, MO USA), nifedipine (cat. no.N7634; Sigma-Aldrich, St. Louis, MO USA), 2-APB (cat. no. D9754; Sigma-Aldrich, St. Louis, MO USA), M871(cat. no. ab141159; Abcam, Cambridge, UK) and SNAP37889 (cat. no. 11L-312S; Key organics, Camelford, UK) were applied separately to block the corresponding pathway. ..

    Recombinant:

    Article Title: Transbilayer phospholipid movement facilitates the translocation of annexin across membranes
    Article Snippet: Antibodies used were: mouse monoclonal anti-annexin A2 (BD Biosciences; 610071; 1:1000), mouse monoclonal anti-annexin A5 (Abcam; ab54775, 1:1000), mouse monoclonal anti-transferrin receptor (Zymed; H68.4; 1:1000), rabbit polyclonal anti-actin (Sigma; A2066; 1:2000), mouse monoclonal anti-tubulin (Sigma; T9026; 1:4000), mouse monoclonal anti-FLAG (Sigma-Aldrich; clone M2; 1:4000), mouse monoclonal anti-Arf1 (Santa Cruz Biotechnology; sc-53168, 1:1000), rat anti-galectin-3 (Biolegend; 125401; western blotting, 1:2000) mouse monoclonal anti-Arf6 (Santa Cruz Biotechnology; clone 3A-1; 1:500), mouse monoclonal anti-LAMP2 (Biolegend; 354302; western blotting, 1:1000), rat polyclonal anti-galectin-3 conjugated to Alexa Fluor 647 (Biolegend; 125408; flow cytometry, 1:100) and rabbit polyclonal anti-mCherry antibody (Genetex; GTX128508-S). .. Reagents used were: cinnamycin (Santa Cruz Biotechnology; sc-391464), mastoparan X (Alfa Aesar; J61173), recombinant annexin-A5–FITC (Abcam; ab14085), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE, Sigma-Aldrich; 54008), 2-oleoyl-1-palmitoyl-sn-glycero-3-phosphocholine (POPC, Sigma-Aldrich; P3017), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(7-nitro-2-1,3-benzoxadiazol-4-yl) (NBD-PE, Avanti Polar Lipids, 810145) and proteinase K (Molecular Biology; BP1700-100), EGTA (Sigma-Aldrich; E3889), sodium dithionite (Sigma-Aldrich; 71699), versene solution containing ethylenediaminetetraacetic acid (EDTA) (Gibco; 15040-033), propidium iodide solution (Biolegend; 421301), QuickExtract DNA extraction solution (Epicenter; QE0905T), Herculase II fusion DNA polymerase (Agilent; 600675), annexin-V–FITC and annexin-V–Cy5 Apoptosis Staining/Detection Kit (ab14085, ab14150), annexin-V conjugated to Alexa Fluor 647 (Biolegend; 640912) and ionomycin (Cayman Chemical Company; 10004974), recombinant galectin-3 (Biolegend; 599706) and recombinant anneaxin A5 (Novus NBP1-30265). .. Oligonucleotides for TMEM16F CRISPR targeting and sequencing were synthesised from Sigma-Aldrich ( ).

    Article Title: Transbilayer phospholipid movement facilitates the translocation of annexin across membranes
    Article Snippet: Antibodies Antibodies used were: mouse monoclonal anti-annexin A2 (BD Biosciences; 610071; 1:1000), mouse monoclonal anti-annexin A5 (Abcam; ab54775, 1:1000), mouse monoclonal anti-transferrin receptor (Zymed; H68.4; 1:1000), rabbit polyclonal anti-actin (Sigma; A2066; 1:2000), mouse monoclonal anti-tubulin (Sigma; T9026; 1:4000), mouse monoclonal anti-FLAG (Sigma-Aldrich; clone M2; 1:4000), mouse monoclonal anti-Arf1 (Santa Cruz Biotechnology; sc-53168, 1:1000), rat anti-galectin-3 (Biolegend; 125401; western blotting, 1:2000) mouse monoclonal anti-Arf6 (Santa Cruz Biotechnology; clone 3A-1; 1:500), mouse monoclonal anti-LAMP2 (Biolegend; 354302; western blotting, 1:1000), rat polyclonal anti-galectin-3 conjugated to Alexa Fluor 647 (Biolegend; 125408; flow cytometry, 1:100) and rabbit polyclonal anti-mCherry antibody (Genetex; GTX128508-S). .. Reagents Reagents used were: cinnamycin (Santa Cruz Biotechnology; sc-391464), mastoparan X (Alfa Aesar; J61173), recombinant annexin-A5–FITC (Abcam; ab14085), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE, Sigma-Aldrich; 54008), 2-oleoyl-1-palmitoyl-sn-glycero-3-phosphocholine (POPC, Sigma-Aldrich; P3017), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(7-nitro-2-1,3-benzoxadiazol-4-yl) (NBD-PE, Avanti Polar Lipids, 810145) and proteinase K (Molecular Biology; BP1700-100), EGTA (Sigma-Aldrich; E3889), sodium dithionite (Sigma-Aldrich; 71699), versene solution containing ethylenediaminetetraacetic acid (EDTA) (Gibco; 15040-033), propidium iodide solution (Biolegend; 421301), QuickExtract DNA extraction solution (Epicenter; QE0905T), Herculase II fusion DNA polymerase (Agilent; 600675), annexin-V–FITC and annexin-V–Cy5 Apoptosis Staining/Detection Kit (ab14085, ab14150), annexin-V conjugated to Alexa Fluor 647 (Biolegend; 640912) and ionomycin (Cayman Chemical Company; 10004974), recombinant galectin-3 (Biolegend; 599706) and recombinant anneaxin A5 (Novus NBP1-30265). .. Oligonucleotides for TMEM16F CRISPR targeting and sequencing were synthesised from Sigma-Aldrich ( Table S1 ).

    DNA Extraction:

    Article Title: Transbilayer phospholipid movement facilitates the translocation of annexin across membranes
    Article Snippet: Antibodies used were: mouse monoclonal anti-annexin A2 (BD Biosciences; 610071; 1:1000), mouse monoclonal anti-annexin A5 (Abcam; ab54775, 1:1000), mouse monoclonal anti-transferrin receptor (Zymed; H68.4; 1:1000), rabbit polyclonal anti-actin (Sigma; A2066; 1:2000), mouse monoclonal anti-tubulin (Sigma; T9026; 1:4000), mouse monoclonal anti-FLAG (Sigma-Aldrich; clone M2; 1:4000), mouse monoclonal anti-Arf1 (Santa Cruz Biotechnology; sc-53168, 1:1000), rat anti-galectin-3 (Biolegend; 125401; western blotting, 1:2000) mouse monoclonal anti-Arf6 (Santa Cruz Biotechnology; clone 3A-1; 1:500), mouse monoclonal anti-LAMP2 (Biolegend; 354302; western blotting, 1:1000), rat polyclonal anti-galectin-3 conjugated to Alexa Fluor 647 (Biolegend; 125408; flow cytometry, 1:100) and rabbit polyclonal anti-mCherry antibody (Genetex; GTX128508-S). .. Reagents used were: cinnamycin (Santa Cruz Biotechnology; sc-391464), mastoparan X (Alfa Aesar; J61173), recombinant annexin-A5–FITC (Abcam; ab14085), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE, Sigma-Aldrich; 54008), 2-oleoyl-1-palmitoyl-sn-glycero-3-phosphocholine (POPC, Sigma-Aldrich; P3017), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(7-nitro-2-1,3-benzoxadiazol-4-yl) (NBD-PE, Avanti Polar Lipids, 810145) and proteinase K (Molecular Biology; BP1700-100), EGTA (Sigma-Aldrich; E3889), sodium dithionite (Sigma-Aldrich; 71699), versene solution containing ethylenediaminetetraacetic acid (EDTA) (Gibco; 15040-033), propidium iodide solution (Biolegend; 421301), QuickExtract DNA extraction solution (Epicenter; QE0905T), Herculase II fusion DNA polymerase (Agilent; 600675), annexin-V–FITC and annexin-V–Cy5 Apoptosis Staining/Detection Kit (ab14085, ab14150), annexin-V conjugated to Alexa Fluor 647 (Biolegend; 640912) and ionomycin (Cayman Chemical Company; 10004974), recombinant galectin-3 (Biolegend; 599706) and recombinant anneaxin A5 (Novus NBP1-30265). .. Oligonucleotides for TMEM16F CRISPR targeting and sequencing were synthesised from Sigma-Aldrich ( ).

    Article Title: Transbilayer phospholipid movement facilitates the translocation of annexin across membranes
    Article Snippet: Antibodies Antibodies used were: mouse monoclonal anti-annexin A2 (BD Biosciences; 610071; 1:1000), mouse monoclonal anti-annexin A5 (Abcam; ab54775, 1:1000), mouse monoclonal anti-transferrin receptor (Zymed; H68.4; 1:1000), rabbit polyclonal anti-actin (Sigma; A2066; 1:2000), mouse monoclonal anti-tubulin (Sigma; T9026; 1:4000), mouse monoclonal anti-FLAG (Sigma-Aldrich; clone M2; 1:4000), mouse monoclonal anti-Arf1 (Santa Cruz Biotechnology; sc-53168, 1:1000), rat anti-galectin-3 (Biolegend; 125401; western blotting, 1:2000) mouse monoclonal anti-Arf6 (Santa Cruz Biotechnology; clone 3A-1; 1:500), mouse monoclonal anti-LAMP2 (Biolegend; 354302; western blotting, 1:1000), rat polyclonal anti-galectin-3 conjugated to Alexa Fluor 647 (Biolegend; 125408; flow cytometry, 1:100) and rabbit polyclonal anti-mCherry antibody (Genetex; GTX128508-S). .. Reagents Reagents used were: cinnamycin (Santa Cruz Biotechnology; sc-391464), mastoparan X (Alfa Aesar; J61173), recombinant annexin-A5–FITC (Abcam; ab14085), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE, Sigma-Aldrich; 54008), 2-oleoyl-1-palmitoyl-sn-glycero-3-phosphocholine (POPC, Sigma-Aldrich; P3017), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(7-nitro-2-1,3-benzoxadiazol-4-yl) (NBD-PE, Avanti Polar Lipids, 810145) and proteinase K (Molecular Biology; BP1700-100), EGTA (Sigma-Aldrich; E3889), sodium dithionite (Sigma-Aldrich; 71699), versene solution containing ethylenediaminetetraacetic acid (EDTA) (Gibco; 15040-033), propidium iodide solution (Biolegend; 421301), QuickExtract DNA extraction solution (Epicenter; QE0905T), Herculase II fusion DNA polymerase (Agilent; 600675), annexin-V–FITC and annexin-V–Cy5 Apoptosis Staining/Detection Kit (ab14085, ab14150), annexin-V conjugated to Alexa Fluor 647 (Biolegend; 640912) and ionomycin (Cayman Chemical Company; 10004974), recombinant galectin-3 (Biolegend; 599706) and recombinant anneaxin A5 (Novus NBP1-30265). .. Oligonucleotides for TMEM16F CRISPR targeting and sequencing were synthesised from Sigma-Aldrich ( Table S1 ).

    Staining:

    Article Title: Transbilayer phospholipid movement facilitates the translocation of annexin across membranes
    Article Snippet: Antibodies used were: mouse monoclonal anti-annexin A2 (BD Biosciences; 610071; 1:1000), mouse monoclonal anti-annexin A5 (Abcam; ab54775, 1:1000), mouse monoclonal anti-transferrin receptor (Zymed; H68.4; 1:1000), rabbit polyclonal anti-actin (Sigma; A2066; 1:2000), mouse monoclonal anti-tubulin (Sigma; T9026; 1:4000), mouse monoclonal anti-FLAG (Sigma-Aldrich; clone M2; 1:4000), mouse monoclonal anti-Arf1 (Santa Cruz Biotechnology; sc-53168, 1:1000), rat anti-galectin-3 (Biolegend; 125401; western blotting, 1:2000) mouse monoclonal anti-Arf6 (Santa Cruz Biotechnology; clone 3A-1; 1:500), mouse monoclonal anti-LAMP2 (Biolegend; 354302; western blotting, 1:1000), rat polyclonal anti-galectin-3 conjugated to Alexa Fluor 647 (Biolegend; 125408; flow cytometry, 1:100) and rabbit polyclonal anti-mCherry antibody (Genetex; GTX128508-S). .. Reagents used were: cinnamycin (Santa Cruz Biotechnology; sc-391464), mastoparan X (Alfa Aesar; J61173), recombinant annexin-A5–FITC (Abcam; ab14085), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE, Sigma-Aldrich; 54008), 2-oleoyl-1-palmitoyl-sn-glycero-3-phosphocholine (POPC, Sigma-Aldrich; P3017), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(7-nitro-2-1,3-benzoxadiazol-4-yl) (NBD-PE, Avanti Polar Lipids, 810145) and proteinase K (Molecular Biology; BP1700-100), EGTA (Sigma-Aldrich; E3889), sodium dithionite (Sigma-Aldrich; 71699), versene solution containing ethylenediaminetetraacetic acid (EDTA) (Gibco; 15040-033), propidium iodide solution (Biolegend; 421301), QuickExtract DNA extraction solution (Epicenter; QE0905T), Herculase II fusion DNA polymerase (Agilent; 600675), annexin-V–FITC and annexin-V–Cy5 Apoptosis Staining/Detection Kit (ab14085, ab14150), annexin-V conjugated to Alexa Fluor 647 (Biolegend; 640912) and ionomycin (Cayman Chemical Company; 10004974), recombinant galectin-3 (Biolegend; 599706) and recombinant anneaxin A5 (Novus NBP1-30265). .. Oligonucleotides for TMEM16F CRISPR targeting and sequencing were synthesised from Sigma-Aldrich ( ).

    Article Title: Transbilayer phospholipid movement facilitates the translocation of annexin across membranes
    Article Snippet: Antibodies Antibodies used were: mouse monoclonal anti-annexin A2 (BD Biosciences; 610071; 1:1000), mouse monoclonal anti-annexin A5 (Abcam; ab54775, 1:1000), mouse monoclonal anti-transferrin receptor (Zymed; H68.4; 1:1000), rabbit polyclonal anti-actin (Sigma; A2066; 1:2000), mouse monoclonal anti-tubulin (Sigma; T9026; 1:4000), mouse monoclonal anti-FLAG (Sigma-Aldrich; clone M2; 1:4000), mouse monoclonal anti-Arf1 (Santa Cruz Biotechnology; sc-53168, 1:1000), rat anti-galectin-3 (Biolegend; 125401; western blotting, 1:2000) mouse monoclonal anti-Arf6 (Santa Cruz Biotechnology; clone 3A-1; 1:500), mouse monoclonal anti-LAMP2 (Biolegend; 354302; western blotting, 1:1000), rat polyclonal anti-galectin-3 conjugated to Alexa Fluor 647 (Biolegend; 125408; flow cytometry, 1:100) and rabbit polyclonal anti-mCherry antibody (Genetex; GTX128508-S). .. Reagents Reagents used were: cinnamycin (Santa Cruz Biotechnology; sc-391464), mastoparan X (Alfa Aesar; J61173), recombinant annexin-A5–FITC (Abcam; ab14085), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE, Sigma-Aldrich; 54008), 2-oleoyl-1-palmitoyl-sn-glycero-3-phosphocholine (POPC, Sigma-Aldrich; P3017), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(7-nitro-2-1,3-benzoxadiazol-4-yl) (NBD-PE, Avanti Polar Lipids, 810145) and proteinase K (Molecular Biology; BP1700-100), EGTA (Sigma-Aldrich; E3889), sodium dithionite (Sigma-Aldrich; 71699), versene solution containing ethylenediaminetetraacetic acid (EDTA) (Gibco; 15040-033), propidium iodide solution (Biolegend; 421301), QuickExtract DNA extraction solution (Epicenter; QE0905T), Herculase II fusion DNA polymerase (Agilent; 600675), annexin-V–FITC and annexin-V–Cy5 Apoptosis Staining/Detection Kit (ab14085, ab14150), annexin-V conjugated to Alexa Fluor 647 (Biolegend; 640912) and ionomycin (Cayman Chemical Company; 10004974), recombinant galectin-3 (Biolegend; 599706) and recombinant anneaxin A5 (Novus NBP1-30265). .. Oligonucleotides for TMEM16F CRISPR targeting and sequencing were synthesised from Sigma-Aldrich ( Table S1 ).

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    Millipore egta
    Cinnamycin facilitates annexin translocation across membranes in liposomes but not galectin-3. (A) Cinnamycin induces lipid movement in liposomes. Left, schematic of the lipid quenching assay. Right, LUVs (PC:PE 1:1, and a trace of <t>NBD-PE)</t> were pre-incubated with cinnamycin (10 µM) or DMSO for 30 min and changes in NBD fluorescence during the experiment time were recorded upon the addition of dithionite and Triton X-100. (B) Cinnamycin increases the <t>EGTA-resistant</t> membrane-bound fraction of annexin A5. PC:PE liposomes (MLVs) in CaCl 2 -containing buffer were incubated with annexin A5. Cinnamycin or DMSO was added for a further 40 min incubation at 37°C. Some of the samples were also treated with EGTA before all samples were centrifuged. Liposome pellets were mixed with boiling SDS sample buffer, separated by SDS-PAGE and analysed by western blotted with anti-annexin A5 antibodies for the detection of the membrane-bound fraction of annexin A5. Results are mean±s.d. comparing lane 3 vs. lane 4; n =3 independent experiments, * P
    Egta, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/egta/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    egta - by Bioz Stars, 2021-03
    99/100 stars
      Buy from Supplier

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    Cinnamycin facilitates annexin translocation across membranes in liposomes but not galectin-3. (A) Cinnamycin induces lipid movement in liposomes. Left, schematic of the lipid quenching assay. Right, LUVs (PC:PE 1:1, and a trace of NBD-PE) were pre-incubated with cinnamycin (10 µM) or DMSO for 30 min and changes in NBD fluorescence during the experiment time were recorded upon the addition of dithionite and Triton X-100. (B) Cinnamycin increases the EGTA-resistant membrane-bound fraction of annexin A5. PC:PE liposomes (MLVs) in CaCl 2 -containing buffer were incubated with annexin A5. Cinnamycin or DMSO was added for a further 40 min incubation at 37°C. Some of the samples were also treated with EGTA before all samples were centrifuged. Liposome pellets were mixed with boiling SDS sample buffer, separated by SDS-PAGE and analysed by western blotted with anti-annexin A5 antibodies for the detection of the membrane-bound fraction of annexin A5. Results are mean±s.d. comparing lane 3 vs. lane 4; n =3 independent experiments, * P

    Journal: Journal of Cell Science

    Article Title: Transbilayer phospholipid movement facilitates the translocation of annexin across membranes

    doi: 10.1242/jcs.217034

    Figure Lengend Snippet: Cinnamycin facilitates annexin translocation across membranes in liposomes but not galectin-3. (A) Cinnamycin induces lipid movement in liposomes. Left, schematic of the lipid quenching assay. Right, LUVs (PC:PE 1:1, and a trace of NBD-PE) were pre-incubated with cinnamycin (10 µM) or DMSO for 30 min and changes in NBD fluorescence during the experiment time were recorded upon the addition of dithionite and Triton X-100. (B) Cinnamycin increases the EGTA-resistant membrane-bound fraction of annexin A5. PC:PE liposomes (MLVs) in CaCl 2 -containing buffer were incubated with annexin A5. Cinnamycin or DMSO was added for a further 40 min incubation at 37°C. Some of the samples were also treated with EGTA before all samples were centrifuged. Liposome pellets were mixed with boiling SDS sample buffer, separated by SDS-PAGE and analysed by western blotted with anti-annexin A5 antibodies for the detection of the membrane-bound fraction of annexin A5. Results are mean±s.d. comparing lane 3 vs. lane 4; n =3 independent experiments, * P

    Article Snippet: Reagents Reagents used were: cinnamycin (Santa Cruz Biotechnology; sc-391464), mastoparan X (Alfa Aesar; J61173), recombinant annexin-A5–FITC (Abcam; ab14085), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE, Sigma-Aldrich; 54008), 2-oleoyl-1-palmitoyl-sn-glycero-3-phosphocholine (POPC, Sigma-Aldrich; P3017), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(7-nitro-2-1,3-benzoxadiazol-4-yl) (NBD-PE, Avanti Polar Lipids, 810145) and proteinase K (Molecular Biology; BP1700-100), EGTA (Sigma-Aldrich; E3889), sodium dithionite (Sigma-Aldrich; 71699), versene solution containing ethylenediaminetetraacetic acid (EDTA) (Gibco; 15040-033), propidium iodide solution (Biolegend; 421301), QuickExtract DNA extraction solution (Epicenter; QE0905T), Herculase II fusion DNA polymerase (Agilent; 600675), annexin-V–FITC and annexin-V–Cy5 Apoptosis Staining/Detection Kit (ab14085, ab14150), annexin-V conjugated to Alexa Fluor 647 (Biolegend; 640912) and ionomycin (Cayman Chemical Company; 10004974), recombinant galectin-3 (Biolegend; 599706) and recombinant anneaxin A5 (Novus NBP1-30265).

    Techniques: Translocation Assay, Incubation, Fluorescence, SDS Page, Western Blot

    A heat-stable, low-molecular weight compound present in CM can induce microcolony dispersal. The CM was pretreated with 95°C heat, a protease inhibitor cocktail, proteinase K, trypsin, chymotrypsin, DNase I, RNase A, EDTA, or EGTA (A) or passaged through a 3 kDa cut-off filter (B) prior to induction assay. Microcolony dispersal was examined in live-cell time-lapse microscopy after addition of treated CM. A black horizontal line in panels represents the 3 h time point when treated CM was added to preformed microcolonies. Data represent the mean ± SD of three independent experiments. *p

    Journal: PLoS Pathogens

    Article Title: Host cell-derived lactate functions as an effector molecule in Neisseria meningitidis microcolony dispersal

    doi: 10.1371/journal.ppat.1006251

    Figure Lengend Snippet: A heat-stable, low-molecular weight compound present in CM can induce microcolony dispersal. The CM was pretreated with 95°C heat, a protease inhibitor cocktail, proteinase K, trypsin, chymotrypsin, DNase I, RNase A, EDTA, or EGTA (A) or passaged through a 3 kDa cut-off filter (B) prior to induction assay. Microcolony dispersal was examined in live-cell time-lapse microscopy after addition of treated CM. A black horizontal line in panels represents the 3 h time point when treated CM was added to preformed microcolonies. Data represent the mean ± SD of three independent experiments. *p

    Article Snippet: EDTA (E5134, Sigma-Aldrich) and EGTA (E4378, Sigma-Aldrich) were used as metal ion chelators at final concentrations of 2 mM.

    Techniques: Molecular Weight, Protease Inhibitor, Time-lapse Microscopy

    Cinnamycin facilitates annexin translocation across membranes in liposomes but not galectin-3. (A) Cinnamycin induces lipid movement in liposomes. Left, schematic of the lipid quenching assay. Right, LUVs (PC:PE 1:1, and a trace of NBD-PE) were pre-incubated with cinnamycin (10 µM) or DMSO for 30 min and changes in NBD fluorescence during the experiment time were recorded upon the addition of dithionite and Triton X-100. (B) Cinnamycin increases the EGTA-resistant membrane-bound fraction of annexin A5. PC:PE liposomes (MLVs) in CaCl 2 -containing buffer were incubated with annexin A5. Cinnamycin or DMSO was added for a further 40 min incubation at 37°C. Some of the samples were also treated with EGTA before all samples were centrifuged. Liposome pellets were mixed with boiling SDS sample buffer, separated by SDS-PAGE and analysed by western blotted with anti-annexin A5 antibodies for the detection of the membrane-bound fraction of annexin A5. Results are mean±s.d. comparing lane 3 vs. lane 4; n =3 independent experiments, * P

    Journal: Journal of Cell Science

    Article Title: Transbilayer phospholipid movement facilitates the translocation of annexin across membranes

    doi: 10.1242/jcs.217034

    Figure Lengend Snippet: Cinnamycin facilitates annexin translocation across membranes in liposomes but not galectin-3. (A) Cinnamycin induces lipid movement in liposomes. Left, schematic of the lipid quenching assay. Right, LUVs (PC:PE 1:1, and a trace of NBD-PE) were pre-incubated with cinnamycin (10 µM) or DMSO for 30 min and changes in NBD fluorescence during the experiment time were recorded upon the addition of dithionite and Triton X-100. (B) Cinnamycin increases the EGTA-resistant membrane-bound fraction of annexin A5. PC:PE liposomes (MLVs) in CaCl 2 -containing buffer were incubated with annexin A5. Cinnamycin or DMSO was added for a further 40 min incubation at 37°C. Some of the samples were also treated with EGTA before all samples were centrifuged. Liposome pellets were mixed with boiling SDS sample buffer, separated by SDS-PAGE and analysed by western blotted with anti-annexin A5 antibodies for the detection of the membrane-bound fraction of annexin A5. Results are mean±s.d. comparing lane 3 vs. lane 4; n =3 independent experiments, * P

    Article Snippet: Reagents used were: cinnamycin (Santa Cruz Biotechnology; sc-391464), mastoparan X (Alfa Aesar; J61173), recombinant annexin-A5–FITC (Abcam; ab14085), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE, Sigma-Aldrich; 54008), 2-oleoyl-1-palmitoyl-sn-glycero-3-phosphocholine (POPC, Sigma-Aldrich; P3017), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(7-nitro-2-1,3-benzoxadiazol-4-yl) (NBD-PE, Avanti Polar Lipids, 810145) and proteinase K (Molecular Biology; BP1700-100), EGTA (Sigma-Aldrich; E3889), sodium dithionite (Sigma-Aldrich; 71699), versene solution containing ethylenediaminetetraacetic acid (EDTA) (Gibco; 15040-033), propidium iodide solution (Biolegend; 421301), QuickExtract DNA extraction solution (Epicenter; QE0905T), Herculase II fusion DNA polymerase (Agilent; 600675), annexin-V–FITC and annexin-V–Cy5 Apoptosis Staining/Detection Kit (ab14085, ab14150), annexin-V conjugated to Alexa Fluor 647 (Biolegend; 640912) and ionomycin (Cayman Chemical Company; 10004974), recombinant galectin-3 (Biolegend; 599706) and recombinant anneaxin A5 (Novus NBP1-30265).

    Techniques: Translocation Assay, Incubation, Fluorescence, SDS Page, Western Blot

    The effect of EDTA (left) and EGTA (right) on barnacle cement polymerization. Unpolymerized cement was incubated with EDTA, EGTA or deionized water for 2 min and then run on SDS-PAGE. Mean intensity (+s.e.m.) for the protein bands labeled in

    Journal: The Journal of Experimental Biology

    Article Title: Barnacle cement: a polymerization model based on evolutionary concepts

    doi: 10.1242/jeb.029884

    Figure Lengend Snippet: The effect of EDTA (left) and EGTA (right) on barnacle cement polymerization. Unpolymerized cement was incubated with EDTA, EGTA or deionized water for 2 min and then run on SDS-PAGE. Mean intensity (+s.e.m.) for the protein bands labeled in

    Article Snippet: EGTA (Sigma-Aldrich no. E4378) and EDTA (Fisher Chemical, no. BP118) were used.

    Techniques: Incubation, SDS Page, Labeling