ethylene glycol tetraacetic acid egta  (Tocris)

 
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  • 96
    Name:
    EGTA
    Description:
    Calcium chelating agent
    Catalog Number:
    2807
    Price:
    None
    Category:
    Biochemicals and Molecular Biology Reagents
    Formula:
    Ethylene glycol-bis(2-aminoethylether)-N,N,N',N'-tetraacetic acid
    Buy from Supplier


    Structured Review

    Tocris ethylene glycol tetraacetic acid egta
    EGTA
    Calcium chelating agent
    https://www.bioz.com/result/ethylene glycol tetraacetic acid egta/product/Tocris
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ethylene glycol tetraacetic acid egta - by Bioz Stars, 2020-07
    96/100 stars

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    Related Articles

    Flow Cytometry:

    Article Title: TMEM203 Is a Novel Regulator of Intracellular Calcium Homeostasis and Is Required for Spermatogenesis
    Article Snippet: .. To measure intracellular store calcium flux cells were treated with 1mM EGTA and subsequently with thapsigargin, ionomycin, or m-3m3FBS (Tocris Bioscience) and changes in fluorescence intensity were monitored on an LSRII flow cytometer (BD Biosciences). .. Values were plotted as the ratio of fluorescence at Ca2+ -bound Indo-1 AM to that of Ca2+ -free Indo-1 AM.

    Fluorescence:

    Article Title: TMEM203 Is a Novel Regulator of Intracellular Calcium Homeostasis and Is Required for Spermatogenesis
    Article Snippet: .. To measure intracellular store calcium flux cells were treated with 1mM EGTA and subsequently with thapsigargin, ionomycin, or m-3m3FBS (Tocris Bioscience) and changes in fluorescence intensity were monitored on an LSRII flow cytometer (BD Biosciences). .. Values were plotted as the ratio of fluorescence at Ca2+ -bound Indo-1 AM to that of Ca2+ -free Indo-1 AM.

    Cytometry:

    Article Title: TMEM203 Is a Novel Regulator of Intracellular Calcium Homeostasis and Is Required for Spermatogenesis
    Article Snippet: .. To measure intracellular store calcium flux cells were treated with 1mM EGTA and subsequently with thapsigargin, ionomycin, or m-3m3FBS (Tocris Bioscience) and changes in fluorescence intensity were monitored on an LSRII flow cytometer (BD Biosciences). .. Values were plotted as the ratio of fluorescence at Ca2+ -bound Indo-1 AM to that of Ca2+ -free Indo-1 AM.

    Purification:

    Article Title: LtpD Is a Novel Legionella pneumophila Effector That Binds Phosphatidylinositol 3-Phosphate and Inositol Monophosphatase IMPA1
    Article Snippet: .. The assay was performed at 37°C for 30 min in assay buffer (50 mM Tris-HCl [pH 8.0]) with 0.7 mM myo -inositol monophosphate, 1 mM EGTA, 2 mM MgCl2 , and 0.5 μg of purified His-IMPA1/ml with or without 0.5 μg of purified GST-LtpD/ml and a 1 mM concentration of IMPA1 inhibitor L-690,330 (Tocris Bioscience). .. The reaction was quenched by the addition of 5 μl of MG acidic solution (Cambridge Biosciences) and incubated for 10 min at room temperature, and then 25 μl of MG blue solution added to visualize released phosphate, followed by incubation for a further 20 min at room temperature.

    Concentration Assay:

    Article Title: LtpD Is a Novel Legionella pneumophila Effector That Binds Phosphatidylinositol 3-Phosphate and Inositol Monophosphatase IMPA1
    Article Snippet: .. The assay was performed at 37°C for 30 min in assay buffer (50 mM Tris-HCl [pH 8.0]) with 0.7 mM myo -inositol monophosphate, 1 mM EGTA, 2 mM MgCl2 , and 0.5 μg of purified His-IMPA1/ml with or without 0.5 μg of purified GST-LtpD/ml and a 1 mM concentration of IMPA1 inhibitor L-690,330 (Tocris Bioscience). .. The reaction was quenched by the addition of 5 μl of MG acidic solution (Cambridge Biosciences) and incubated for 10 min at room temperature, and then 25 μl of MG blue solution added to visualize released phosphate, followed by incubation for a further 20 min at room temperature.

    Incubation:

    Article Title: Pak1 Kinase Promotes Activated T Cell Trafficking by Regulating the Expression of L-Selectin and CCR7
    Article Snippet: .. Where indicated, CD4+ T blast cells were incubated for 20 min with DMSO (1:1,000), 10 μM BAPTA-AM (Invitrogen) and 2 mM EGTA or 0.5 μM of the ADAM17 inhibitor TMI1 (Tocris Bioscience). ..

    other:

    Article Title: Nociceptin Signaling Involves a Calcium-Based Depolarization in Tetrahymena thermophila
    Article Snippet: All nociceptin isoforms, thapsigargin, J-113397, and EGTA, were purchased from Tocris Biosciences, Bristol, UK.

    Article Title: On the Mechanism of Synaptic Depression Induced by CaMKIIN, an Endogenous Inhibitor of CaMKII
    Article Snippet: N-methyl-D-aspartic acid (NMDA) was obtained from Sigma Aldrich, and Anisomycin, EGTA, kynurenic acid, thapsigargin and MG 132 were from Tocris Bioscience.

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  • egta  (Tocris)
    96
    Tocris egta
    Protective effects of PMC-12 on cellular Ca 2+ -related ROS production in HT22 cells. Cells were pretreated with 10 μg/ml of PMC-12 for 24 h, followed by exposure to 5 mM glutamate for 24 h. Cells were treated with the specific Ca 2+ inhibitors, 1.5 mM <t>EGTA</t> or 10 μM <t>BAPTA-AM,</t> for 30 min before addition of PMC-12 or glutamate. ROS production was measured using a fluorescence plate reader and mean fluorescence intensity was expressed as the mean ± SEM of three independent experiments. ### P
    Egta, supplied by Tocris, used in various techniques. Bioz Stars score: 96/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/egta/product/Tocris
    Average 96 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    egta - by Bioz Stars, 2020-07
    96/100 stars
      Buy from Supplier

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    Protective effects of PMC-12 on cellular Ca 2+ -related ROS production in HT22 cells. Cells were pretreated with 10 μg/ml of PMC-12 for 24 h, followed by exposure to 5 mM glutamate for 24 h. Cells were treated with the specific Ca 2+ inhibitors, 1.5 mM EGTA or 10 μM BAPTA-AM, for 30 min before addition of PMC-12 or glutamate. ROS production was measured using a fluorescence plate reader and mean fluorescence intensity was expressed as the mean ± SEM of three independent experiments. ### P

    Journal: BMC Complementary and Alternative Medicine

    Article Title: Neuroprotection and spatial memory enhancement of four herbal mixture extract in HT22 hippocampal cells and a mouse model of focal cerebral ischemia

    doi: 10.1186/s12906-015-0741-1

    Figure Lengend Snippet: Protective effects of PMC-12 on cellular Ca 2+ -related ROS production in HT22 cells. Cells were pretreated with 10 μg/ml of PMC-12 for 24 h, followed by exposure to 5 mM glutamate for 24 h. Cells were treated with the specific Ca 2+ inhibitors, 1.5 mM EGTA or 10 μM BAPTA-AM, for 30 min before addition of PMC-12 or glutamate. ROS production was measured using a fluorescence plate reader and mean fluorescence intensity was expressed as the mean ± SEM of three independent experiments. ### P

    Article Snippet: BAPTA-AM and EGTA were purchased from Tocris Bioscience (Ellisville, MO, USA).

    Techniques: Fluorescence

    Altered calcium homeostasis in Tmem203 deficient Mouse Embryonic Fibroblast cells. (A) Cytoplasmic calcium flux were measured by flow cytometry from MEF cells derived from Tmem203—WT (Blue), HET (Green) and null (Red) mice. Treatment with 1 μM TG and EGTA in calcium free buffer showing ER-released calcium flux. Data are representative of at least 3 experiments from multiple MEF derived from littermates. (B) As described in (A), the calcium flux were measured upon treatment with 50 μM m-3M3FBS. (C) As described in (A), the calcium flux were measured upon treatment with 5 μM Ionomycin. (D) Single cell fluorescent microscopy based direct ER calcium measurements in D1ER-HEK293 cells transfected with non-targeting or TMEM203 specific siRNA. Basal ER calcium levels and ER calcium release upon TG treatment was monitored in siRNA or control siRNA transfected D1ER-HEK293 cells. The measurements showed reduced calcium levels in TMEM203 knock down cells but similar TG induced calcium leak kinetics. [Mean; +/- SE; n = 47 (non targeting siRNA) n = 54 (TMEM203 siRNA)].

    Journal: PLoS ONE

    Article Title: TMEM203 Is a Novel Regulator of Intracellular Calcium Homeostasis and Is Required for Spermatogenesis

    doi: 10.1371/journal.pone.0127480

    Figure Lengend Snippet: Altered calcium homeostasis in Tmem203 deficient Mouse Embryonic Fibroblast cells. (A) Cytoplasmic calcium flux were measured by flow cytometry from MEF cells derived from Tmem203—WT (Blue), HET (Green) and null (Red) mice. Treatment with 1 μM TG and EGTA in calcium free buffer showing ER-released calcium flux. Data are representative of at least 3 experiments from multiple MEF derived from littermates. (B) As described in (A), the calcium flux were measured upon treatment with 50 μM m-3M3FBS. (C) As described in (A), the calcium flux were measured upon treatment with 5 μM Ionomycin. (D) Single cell fluorescent microscopy based direct ER calcium measurements in D1ER-HEK293 cells transfected with non-targeting or TMEM203 specific siRNA. Basal ER calcium levels and ER calcium release upon TG treatment was monitored in siRNA or control siRNA transfected D1ER-HEK293 cells. The measurements showed reduced calcium levels in TMEM203 knock down cells but similar TG induced calcium leak kinetics. [Mean; +/- SE; n = 47 (non targeting siRNA) n = 54 (TMEM203 siRNA)].

    Article Snippet: To measure intracellular store calcium flux cells were treated with 1mM EGTA and subsequently with thapsigargin, ionomycin, or m-3m3FBS (Tocris Bioscience) and changes in fluorescence intensity were monitored on an LSRII flow cytometer (BD Biosciences).

    Techniques: Flow Cytometry, Cytometry, Derivative Assay, Mouse Assay, Microscopy, Transfection

    Intracellular store calcium flux and store operated calcium entry kinetics in testicular cells from WT and Tmem203 null mice. Flou3 and Fura red loaded testicular cells prepared from WT and Tmem203 null mice were analyzed by flow cytometry to follow cytosolic calcium kinetics in the gated predominately round spermatids population. Intracellular store calcium flux was measured by recording Flou3 calcium bound to Fura red calcium free ratio in the presence of 1mM EGTA in response to SERCA inhibitor- Thapsigargin (A); Calcium ionophore—Ionomycin (B); Similarly the store operated calcium entry kinetics was followed in WT and Tmem203 null testicular round spermatids gated population by depleting the stores by Thapsigargin (C) or Ionomycin (D) followed with addition of 2mM CaCl 2 .

    Journal: PLoS ONE

    Article Title: TMEM203 Is a Novel Regulator of Intracellular Calcium Homeostasis and Is Required for Spermatogenesis

    doi: 10.1371/journal.pone.0127480

    Figure Lengend Snippet: Intracellular store calcium flux and store operated calcium entry kinetics in testicular cells from WT and Tmem203 null mice. Flou3 and Fura red loaded testicular cells prepared from WT and Tmem203 null mice were analyzed by flow cytometry to follow cytosolic calcium kinetics in the gated predominately round spermatids population. Intracellular store calcium flux was measured by recording Flou3 calcium bound to Fura red calcium free ratio in the presence of 1mM EGTA in response to SERCA inhibitor- Thapsigargin (A); Calcium ionophore—Ionomycin (B); Similarly the store operated calcium entry kinetics was followed in WT and Tmem203 null testicular round spermatids gated population by depleting the stores by Thapsigargin (C) or Ionomycin (D) followed with addition of 2mM CaCl 2 .

    Article Snippet: To measure intracellular store calcium flux cells were treated with 1mM EGTA and subsequently with thapsigargin, ionomycin, or m-3m3FBS (Tocris Bioscience) and changes in fluorescence intensity were monitored on an LSRII flow cytometer (BD Biosciences).

    Techniques: Mouse Assay, Flow Cytometry, Cytometry

    TMEM203 interacts with regulators of ER calcium stores and overexpression depletes ER calcium stores. (A) Confocal analysis of HeLa cells transiently expressing TMEM203-GFP with organelle specific markers for ER (top:Calreticulin-RFP), Mitochondria (middle:BDHA-RFP) or plasma membrane (bottom:LCK-RFP). Separation or colocalization of TMEM203-GFP and organelle marker(s) were visualized by the linescan function of MetaMorph: the fluorescence intensity of each pixel of the line of interest (white lines ~ 75 μm) is shown as a xy-graph for the corresponding green and red channels. The line scan shows that TMEM203-GFP predominately overlapped with the ER marker. (Representative of ~ 50 cells from 2 independent experiments). Note, we cannot rule out that TMEM203 is completely absent from the the mitochondria. (B) Western analysis of complexes immune-precipitated TMEM203-Flag from HEK293 cells with indicated antibodies shows specific interaction with endogenous STIM1, IP3R and SERCA2. (Representative of atleast 2 independent experiments). (C) pTUNE-TMEM203-293cells were treated with the indicated dose of IPTG for 48 hrs to induce TMEM203 expression. Levels of TMEM203-Flag protein were detected by western blot. (D) These IPTG induced cells were subjected to Indo-1 based calcium flux measurements by flow cytometry by first treating with thapsigargin (TG) and EGTA. (E) As in (D) but the cells were treated with Ionomycin. (F) As in (D) but following TG treatment CaCl 2 was added to record SOCE.

    Journal: PLoS ONE

    Article Title: TMEM203 Is a Novel Regulator of Intracellular Calcium Homeostasis and Is Required for Spermatogenesis

    doi: 10.1371/journal.pone.0127480

    Figure Lengend Snippet: TMEM203 interacts with regulators of ER calcium stores and overexpression depletes ER calcium stores. (A) Confocal analysis of HeLa cells transiently expressing TMEM203-GFP with organelle specific markers for ER (top:Calreticulin-RFP), Mitochondria (middle:BDHA-RFP) or plasma membrane (bottom:LCK-RFP). Separation or colocalization of TMEM203-GFP and organelle marker(s) were visualized by the linescan function of MetaMorph: the fluorescence intensity of each pixel of the line of interest (white lines ~ 75 μm) is shown as a xy-graph for the corresponding green and red channels. The line scan shows that TMEM203-GFP predominately overlapped with the ER marker. (Representative of ~ 50 cells from 2 independent experiments). Note, we cannot rule out that TMEM203 is completely absent from the the mitochondria. (B) Western analysis of complexes immune-precipitated TMEM203-Flag from HEK293 cells with indicated antibodies shows specific interaction with endogenous STIM1, IP3R and SERCA2. (Representative of atleast 2 independent experiments). (C) pTUNE-TMEM203-293cells were treated with the indicated dose of IPTG for 48 hrs to induce TMEM203 expression. Levels of TMEM203-Flag protein were detected by western blot. (D) These IPTG induced cells were subjected to Indo-1 based calcium flux measurements by flow cytometry by first treating with thapsigargin (TG) and EGTA. (E) As in (D) but the cells were treated with Ionomycin. (F) As in (D) but following TG treatment CaCl 2 was added to record SOCE.

    Article Snippet: To measure intracellular store calcium flux cells were treated with 1mM EGTA and subsequently with thapsigargin, ionomycin, or m-3m3FBS (Tocris Bioscience) and changes in fluorescence intensity were monitored on an LSRII flow cytometer (BD Biosciences).

    Techniques: Over Expression, Expressing, Marker, Fluorescence, Western Blot, Flow Cytometry, Cytometry

    Pak1 regulates L-selectin shedding in activated CD4 + T cells. (A) Analysis by ELISA of soluble L-selectin in the supernatants of WT and Pak1 (T) −/− T cell blasts. Data were pooled from six independent experiments. (B) Representative time-dependent calcium flux (indo-violet/indo-blue) of Indo-1AM loaded WT and Pak1 (T) −/− CD4 + T cell blasts, stimulated with CCL21 (200 ng/mL) for the indicated time. Results from one of three independent experiments are shown. (C) Calmodulin (CaM) was immunoprecipitated and eluted material was blotted for co-precipitation L-selectin and CaM in lysates prepared from WT and Pak1 (T) −/− T cell blasts. Whole-cell lysates were blotted for CaM. Results from two independent experiments are shown. (D) Analysis of soluble L-selectin in the supernatant of WT and Pak1 (T) −/− CD4 + T cell blasts pre-treated for 20 min with DMSO (1:1,000), BAPTA-AM 10 μM and EGTA 2 mM, or TMI1 (0.5 μM), and incubated for 30 min in the presence of 100 ng/mL CCL21. (E) Left, L-selectin surface marker expression as measured by flow cytometry from WT and Pak1 (T) −/− CD4 + T cell blasts cultured with or without TMI1(10 μM) for 6 days. The results are representative of four experiments. Right, bar chart shows MFI (%) of L-selectin in WT and Pak1 (T) −/− blast T cells. (F) Percentage of WT, Pak1 (T) −/− and Pak1 (T) −/− treated with TMI1 cells per field of view. Statistical analysis: unpaired Student's t -test (A) ; ANOVA between selected columns with Sidak's multiple comparison test (D) ; ANOVA with Tukey's multiple comparison test (E,F) . ns , not significant; * P

    Journal: Frontiers in Immunology

    Article Title: Pak1 Kinase Promotes Activated T Cell Trafficking by Regulating the Expression of L-Selectin and CCR7

    doi: 10.3389/fimmu.2019.00370

    Figure Lengend Snippet: Pak1 regulates L-selectin shedding in activated CD4 + T cells. (A) Analysis by ELISA of soluble L-selectin in the supernatants of WT and Pak1 (T) −/− T cell blasts. Data were pooled from six independent experiments. (B) Representative time-dependent calcium flux (indo-violet/indo-blue) of Indo-1AM loaded WT and Pak1 (T) −/− CD4 + T cell blasts, stimulated with CCL21 (200 ng/mL) for the indicated time. Results from one of three independent experiments are shown. (C) Calmodulin (CaM) was immunoprecipitated and eluted material was blotted for co-precipitation L-selectin and CaM in lysates prepared from WT and Pak1 (T) −/− T cell blasts. Whole-cell lysates were blotted for CaM. Results from two independent experiments are shown. (D) Analysis of soluble L-selectin in the supernatant of WT and Pak1 (T) −/− CD4 + T cell blasts pre-treated for 20 min with DMSO (1:1,000), BAPTA-AM 10 μM and EGTA 2 mM, or TMI1 (0.5 μM), and incubated for 30 min in the presence of 100 ng/mL CCL21. (E) Left, L-selectin surface marker expression as measured by flow cytometry from WT and Pak1 (T) −/− CD4 + T cell blasts cultured with or without TMI1(10 μM) for 6 days. The results are representative of four experiments. Right, bar chart shows MFI (%) of L-selectin in WT and Pak1 (T) −/− blast T cells. (F) Percentage of WT, Pak1 (T) −/− and Pak1 (T) −/− treated with TMI1 cells per field of view. Statistical analysis: unpaired Student's t -test (A) ; ANOVA between selected columns with Sidak's multiple comparison test (D) ; ANOVA with Tukey's multiple comparison test (E,F) . ns , not significant; * P

    Article Snippet: Where indicated, CD4+ T blast cells were incubated for 20 min with DMSO (1:1,000), 10 μM BAPTA-AM (Invitrogen) and 2 mM EGTA or 0.5 μM of the ADAM17 inhibitor TMI1 (Tocris Bioscience).

    Techniques: Enzyme-linked Immunosorbent Assay, Chick Chorioallantoic Membrane Assay, Immunoprecipitation, Incubation, Marker, Expressing, Flow Cytometry, Cytometry, Cell Culture