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Millipore m egta
M Egta, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 112 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/m egta/product/Millipore
Average 92 stars, based on 112 article reviews
Price from $9.99 to $1999.99
m egta - by Bioz Stars, 2020-09
92/100 stars

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Binding Assay:

Article Title: Recruitment of Nox4 to a Plasma Membrane Scaffold Is Required for Localized Reactive Oxygen Species Generation and Sustained Src Activation in Response to Insulin-like Growth Factor-I *
Article Snippet: .. For Nox4/Grb2 or SHP-2 binding assays, T n T expressed Nox4 WT or Nox4 Y491F (100 ng each) was enriched by immunoprecipitating with an anti-Nox4 antibody and phosphorylated by active Src (150 unit) in a buffer containing 50 m m HEPES-NaOH (pH 7.6), 3 m m MnCl2 , 10 m m MgCl2 , 0.1 m m EGTA, 1 m m dithiothreitol, 0.1 m m Na3 O4 , and 0.2 m m ATP and incubating at 30 °C for 30 min. After extensive washing with the above buffer, purified SHP2 (Millipore, Billerica, MA; 500 ng) or Grb2 (Santa Cruz Biotechnology, Inc., Santa Cruz, CA; 500 ng) was added and incubated for 2 h at 4 °C in the binding buffer (500 μl). .. After extensive washing with the binding buffer, the precipitated proteins were separated using 10% SDS-PAGE and immunoblotted with an anti-Grb2 or SHP2 antibody, respectively.

Homogenization:

Article Title: Exercise-induced liver chemokine CXCL-1 expression is linked to muscle-derived interleukin-6 expression
Article Snippet: .. Total protein from muscle and liver was extracted in 500 μl homogenisation buffer (50 m m Tris-HCl (pH 7.4), 150 m m NaCl, 1 m m EGTA, 1 m m EDTA, 0.25% sodium deoxycholate, 1% Triton X-100) containing sodium orthovanadate, phosphatase inhibitor cocktails 1 and 2 (Sigma-Aldrich) and a complete protease inhibitor cocktail (Roche Basel, Switzerland), followed by homogenisation in pre-cooled racks using a Tissuelyzer (Qiagen, Valencia, CA, USA) for 1 min at 30 Hz followed by 15 min incubation on ice. .. The supernatant protein concentrations were determined using a Bio-Rad DC kit (Bio-Rad, Hercules, CA, USA) and bovine serum albumin as standard.

Protease Inhibitor:

Article Title: Exercise-induced liver chemokine CXCL-1 expression is linked to muscle-derived interleukin-6 expression
Article Snippet: .. Total protein from muscle and liver was extracted in 500 μl homogenisation buffer (50 m m Tris-HCl (pH 7.4), 150 m m NaCl, 1 m m EGTA, 1 m m EDTA, 0.25% sodium deoxycholate, 1% Triton X-100) containing sodium orthovanadate, phosphatase inhibitor cocktails 1 and 2 (Sigma-Aldrich) and a complete protease inhibitor cocktail (Roche Basel, Switzerland), followed by homogenisation in pre-cooled racks using a Tissuelyzer (Qiagen, Valencia, CA, USA) for 1 min at 30 Hz followed by 15 min incubation on ice. .. The supernatant protein concentrations were determined using a Bio-Rad DC kit (Bio-Rad, Hercules, CA, USA) and bovine serum albumin as standard.

Article Title: Toll-like receptor 2 deficiency hyperactivates the FoxO1 transcription factor and induces aging-associated cardiac dysfunction in mice
Article Snippet: .. Mice heart tissue and cell lysates were prepared with lysis buffer containing 1 m m EDTA, 1 m m EGTA, 50 m m Tris-HCl, pH 7.8, 1% Nonidet P-40, 150 m m NaCl, 0.5% sodium deoxycholate, and 1% SDS supplemented with sodium orthovanadate, sodium pyrophosphate, a protease inhibitor mixture (Sigma), and phenylmethylsulfonyl fluoride. ..

Article Title: Chronic Antidepressants Reduce Depolarization-Evoked Glutamate Release and Protein Interactions Favoring Formation of SNARE Complex in Hippocampus
Article Snippet: .. For all of the other experiments, synaptosomes were resuspended in lysis buffer: 120 m m NaCl, 20 m m HEPES, pH 7.4, 0.1 m m EGTA, 0.1 m m DTT, containing 20 m m NaF, 5 m m Na4 P2 O7 , 1 m m Na2 VO4 , and 2 μl/ml protease inhibitor mixture (Sigma, St. Louis, MO). .. Additional fractionation of purified synaptosomes into synaptic membrane fraction (LP1) and synaptic vesicle fraction (LP2) was performed by differential centrifugation and ultracentrifugation as reported previously ( ).

Article Title: Impairment of Transforming Growth Factor ? Signaling in Caveolin-1-deficient Hepatocytes
Article Snippet: .. Samples were homogenized in a medium containing 10 m m Tris/HCl, pH 7.5, 1 m m MgCl2 , 1 m m EGTA, 10% (v/v) glycerol, 0.5% (w/v) CHAPS, 1 m m β-mercaptoethanol, 0.1 m m phenylmethylsulfonyl fluoride, Protease Inhibitor Mixture (Sigma) and Phosphatase Inhibitor Mixture 1 and 2 (Sigma). ..

Purification:

Article Title: Recruitment of Nox4 to a Plasma Membrane Scaffold Is Required for Localized Reactive Oxygen Species Generation and Sustained Src Activation in Response to Insulin-like Growth Factor-I *
Article Snippet: .. For Nox4/Grb2 or SHP-2 binding assays, T n T expressed Nox4 WT or Nox4 Y491F (100 ng each) was enriched by immunoprecipitating with an anti-Nox4 antibody and phosphorylated by active Src (150 unit) in a buffer containing 50 m m HEPES-NaOH (pH 7.6), 3 m m MnCl2 , 10 m m MgCl2 , 0.1 m m EGTA, 1 m m dithiothreitol, 0.1 m m Na3 O4 , and 0.2 m m ATP and incubating at 30 °C for 30 min. After extensive washing with the above buffer, purified SHP2 (Millipore, Billerica, MA; 500 ng) or Grb2 (Santa Cruz Biotechnology, Inc., Santa Cruz, CA; 500 ng) was added and incubated for 2 h at 4 °C in the binding buffer (500 μl). .. After extensive washing with the binding buffer, the precipitated proteins were separated using 10% SDS-PAGE and immunoblotted with an anti-Grb2 or SHP2 antibody, respectively.

Incubation:

Article Title: Recruitment of Nox4 to a Plasma Membrane Scaffold Is Required for Localized Reactive Oxygen Species Generation and Sustained Src Activation in Response to Insulin-like Growth Factor-I *
Article Snippet: .. For Nox4/Grb2 or SHP-2 binding assays, T n T expressed Nox4 WT or Nox4 Y491F (100 ng each) was enriched by immunoprecipitating with an anti-Nox4 antibody and phosphorylated by active Src (150 unit) in a buffer containing 50 m m HEPES-NaOH (pH 7.6), 3 m m MnCl2 , 10 m m MgCl2 , 0.1 m m EGTA, 1 m m dithiothreitol, 0.1 m m Na3 O4 , and 0.2 m m ATP and incubating at 30 °C for 30 min. After extensive washing with the above buffer, purified SHP2 (Millipore, Billerica, MA; 500 ng) or Grb2 (Santa Cruz Biotechnology, Inc., Santa Cruz, CA; 500 ng) was added and incubated for 2 h at 4 °C in the binding buffer (500 μl). .. After extensive washing with the binding buffer, the precipitated proteins were separated using 10% SDS-PAGE and immunoblotted with an anti-Grb2 or SHP2 antibody, respectively.

Article Title: Exercise-induced liver chemokine CXCL-1 expression is linked to muscle-derived interleukin-6 expression
Article Snippet: .. Total protein from muscle and liver was extracted in 500 μl homogenisation buffer (50 m m Tris-HCl (pH 7.4), 150 m m NaCl, 1 m m EGTA, 1 m m EDTA, 0.25% sodium deoxycholate, 1% Triton X-100) containing sodium orthovanadate, phosphatase inhibitor cocktails 1 and 2 (Sigma-Aldrich) and a complete protease inhibitor cocktail (Roche Basel, Switzerland), followed by homogenisation in pre-cooled racks using a Tissuelyzer (Qiagen, Valencia, CA, USA) for 1 min at 30 Hz followed by 15 min incubation on ice. .. The supernatant protein concentrations were determined using a Bio-Rad DC kit (Bio-Rad, Hercules, CA, USA) and bovine serum albumin as standard.

Activity Assay:

Article Title: Cross-phosphorylation between Arabidopsis thaliana
Article Snippet: .. Kinase activity was measured in the presence of 2 μCi of [γ-32 P]ATP (500 μCi/mmol) and 90 μ m AMARA peptide (AMARAASAAALARRR) ( ) in kinase activity buffer containing 0.1 m HEPES-NaOH, pH 7.3; 5 m m dithiothreitol; 10 m m MgCl2 ; 0.5 m m EGTA; 20, 30, or 1000 μ m ATP; 1 μl of antiprotease mixture (P9599; Sigma-Aldrich) for 1 ml of buffer; 1 μl of each antiphosphatase mixture (P2850 and P5726; Sigma-Aldrich) for 1 ml of buffer, at 30 °C in a total volume of 50 μl. .. Following 30–60 min of incubation, an aliquot of 10 μl (two replicates) was spotted onto 1 cm2 of Whatman P81 cation-exchange paper.

Isolation:

Article Title: Golgi- and Trans-Golgi Network-Mediated Vesicle Trafficking Is Required for Wax Secretion from Epidermal Cells
Article Snippet: .. Microsomes were isolated by homogenizing seedlings in 50 m m HEPES buffer (pH 7.6) with 400 m m Suc, 10 m m KCl, 3 m m EGTA, 3 m m Na-EDTA, 0.1% (w/v) fatty acid-free bovine serum albumin (Sigma), 1.5 m m dithiothreitol, and 1 m m sodium ascorbate, filtrating the homogenate through Miracloth (Calbiochem), centrifuging out large contaminants (6,000 g for 15 min at 4°C), and centrifuging the supernatant at 60,000 g for 30 min at 4°C. ..

Lysis:

Article Title: Toll-like receptor 2 deficiency hyperactivates the FoxO1 transcription factor and induces aging-associated cardiac dysfunction in mice
Article Snippet: .. Mice heart tissue and cell lysates were prepared with lysis buffer containing 1 m m EDTA, 1 m m EGTA, 50 m m Tris-HCl, pH 7.8, 1% Nonidet P-40, 150 m m NaCl, 0.5% sodium deoxycholate, and 1% SDS supplemented with sodium orthovanadate, sodium pyrophosphate, a protease inhibitor mixture (Sigma), and phenylmethylsulfonyl fluoride. ..

Article Title: Chronic Antidepressants Reduce Depolarization-Evoked Glutamate Release and Protein Interactions Favoring Formation of SNARE Complex in Hippocampus
Article Snippet: .. For all of the other experiments, synaptosomes were resuspended in lysis buffer: 120 m m NaCl, 20 m m HEPES, pH 7.4, 0.1 m m EGTA, 0.1 m m DTT, containing 20 m m NaF, 5 m m Na4 P2 O7 , 1 m m Na2 VO4 , and 2 μl/ml protease inhibitor mixture (Sigma, St. Louis, MO). .. Additional fractionation of purified synaptosomes into synaptic membrane fraction (LP1) and synaptic vesicle fraction (LP2) was performed by differential centrifugation and ultracentrifugation as reported previously ( ).

Mouse Assay:

Article Title: Toll-like receptor 2 deficiency hyperactivates the FoxO1 transcription factor and induces aging-associated cardiac dysfunction in mice
Article Snippet: .. Mice heart tissue and cell lysates were prepared with lysis buffer containing 1 m m EDTA, 1 m m EGTA, 50 m m Tris-HCl, pH 7.8, 1% Nonidet P-40, 150 m m NaCl, 0.5% sodium deoxycholate, and 1% SDS supplemented with sodium orthovanadate, sodium pyrophosphate, a protease inhibitor mixture (Sigma), and phenylmethylsulfonyl fluoride. ..

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    Millipore m egta
    Lysophosphatidic acid (LPA) and sphingosine 1-phosphate (S1P) induce Ca 2+ mobilization and phospholipase D (PLD) activation in type II alveolar epithelial cells. (a, c) A549 cells were labelled with 3 μ m Fluo-3/AM, treated or not with <t>EDTA,</t> <t>EGTA</t>
    M Egta, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 112 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/m egta/product/Millipore
    Average 92 stars, based on 112 article reviews
    Price from $9.99 to $1999.99
    m egta - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

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    Lysophosphatidic acid (LPA) and sphingosine 1-phosphate (S1P) induce Ca 2+ mobilization and phospholipase D (PLD) activation in type II alveolar epithelial cells. (a, c) A549 cells were labelled with 3 μ m Fluo-3/AM, treated or not with EDTA, EGTA

    Journal: Immunology

    Article Title: Natural lysophospholipids reduce Mycobacterium tuberculosis-induced cytotoxicity and induce anti-mycobacterial activity by a phagolysosome maturation-dependent mechanism in A549 type II alveolar epithelial cells

    doi: 10.1111/j.1365-2567.2009.03145.x

    Figure Lengend Snippet: Lysophosphatidic acid (LPA) and sphingosine 1-phosphate (S1P) induce Ca 2+ mobilization and phospholipase D (PLD) activation in type II alveolar epithelial cells. (a, c) A549 cells were labelled with 3 μ m Fluo-3/AM, treated or not with EDTA, EGTA

    Article Snippet: In several experiments, 2 m m EDTA or 3 m m EGTA (Calbiochem) were added 15 min before the addition of LPA or S1P, whereas 20 μ m 1,2-bis(2-aminophenoxy)ethane-N,N,N1,N1-tetraacetic acid tetraicis (acetoxymethyl ester) (BAPTA-AM) (Sigma-Aldrich, Milan, Italy) was added 30 min before stimulation with LPA or S1P.

    Techniques: Activation Assay