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Millipore egta am
Egta Am, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/egta am/product/Millipore
Average 97 stars, based on 1 article reviews
Price from $9.99 to $1999.99
egta am - by Bioz Stars, 2021-03
97/100 stars

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In Vitro:

Article Title: Glutamate dehydrogenase 1 signals through antioxidant glutathione peroxidase 1 to regulate redox homeostasis and tumor growth
Article Snippet: Tumor proliferation was determined by Ki-67 IHC staining. .. To screen potential GDH1 inhibitors, in vitro GDH1 activity assay was performed as described above in presence of compounds (10 µM) using GDH1-overexpressing 293T cell lysates or purified GDH (Sigma). ..

Article Title: A p53/lnc‐Ip53 Negative Feedback Loop Regulates Tumor Growth and Chemoresistance, A p53/lnc‐Ip53 Negative Feedback Loop Regulates Tumor Growth and Chemoresistance
Article Snippet: Protein acetylation assay was then performed as reported[ ] with modifications. .. Ten µL of affinity gel‐bound Flag, Flag‐p300 or Flag‐p300‐3 were preincubated with 1 µg of in vitro transcribed unlabeled lnc‐Ip53 or lnc‐Ip53‐AS at RT with shaking (800 r.p.m.) for 30 min, followed by adding 1 µg of purified GST‐p53, 0.5 mm acetyl coenzyme A (Ac‐CoA; A2056; Sigma‐Aldrich, St Louis, MO, USA), 2 × HAT buffer and DEPC‐H2O to make up a final volume of 30 µL reaction mixture containing 1 × HAT buffer, then incubated at 30 °C with shaking (800 r.p.m.) for 90 min before immunoblotting. .. Cell Cycle AnalysisCells were stained with propidium iodide (PI), then analyzed by fluorescence‐activated cell sorting (FACS; Gallios, Beckman Coulter, Miami, FL, USA).

Article Title: Volumetric muscle loss injury repair using in situ fibrin gel cast seeded with muscle-derived stem cells (MDSCs)
Article Snippet: MDSCs were expanded from PP6 and labeled with a membrane LacZ lentivirus for in vivo experiments. .. For in vitro cell/gel casting, MDSCs (1 × 106 ) were suspended in 200 μl of fibrinogen solution (4mg/ml, Sigma, F4753) and gel formation was induced by addition of thrombin (5IU, Sigma, T4648) in a 48 well plate. ..

Article Title: S-nitrosylated SHP-2 contributes to NMDA receptor-mediated excitotoxicity in acute ischemic stroke
Article Snippet: For the biotin-switch assay, cortical hemispheres ( n = 3) were harvested from the control or ischemic side of the brain immediately after reperfusion. .. In vitro phosphatase activity of recombinant Shp-2 with pNPP (Sigma) as a substrate was measured in a 30-μL reaction mixture containing 20 mM pNPP, 50 mM sodium citrate (pH 4.8), and 1 μM recombinant SHP-2 ( ). .. After 30 min incubation at 37 °C, the reaction was terminated by addition of 1 M NaOH (270 μL).

Activity Assay:

Article Title: Glutamate dehydrogenase 1 signals through antioxidant glutathione peroxidase 1 to regulate redox homeostasis and tumor growth
Article Snippet: Tumor proliferation was determined by Ki-67 IHC staining. .. To screen potential GDH1 inhibitors, in vitro GDH1 activity assay was performed as described above in presence of compounds (10 µM) using GDH1-overexpressing 293T cell lysates or purified GDH (Sigma). ..

Article Title: S-nitrosylated SHP-2 contributes to NMDA receptor-mediated excitotoxicity in acute ischemic stroke
Article Snippet: For the biotin-switch assay, cortical hemispheres ( n = 3) were harvested from the control or ischemic side of the brain immediately after reperfusion. .. In vitro phosphatase activity of recombinant Shp-2 with pNPP (Sigma) as a substrate was measured in a 30-μL reaction mixture containing 20 mM pNPP, 50 mM sodium citrate (pH 4.8), and 1 μM recombinant SHP-2 ( ). .. After 30 min incubation at 37 °C, the reaction was terminated by addition of 1 M NaOH (270 μL).

Purification:

Article Title: Glutamate dehydrogenase 1 signals through antioxidant glutathione peroxidase 1 to regulate redox homeostasis and tumor growth
Article Snippet: Tumor proliferation was determined by Ki-67 IHC staining. .. To screen potential GDH1 inhibitors, in vitro GDH1 activity assay was performed as described above in presence of compounds (10 µM) using GDH1-overexpressing 293T cell lysates or purified GDH (Sigma). ..

Article Title: A p53/lnc‐Ip53 Negative Feedback Loop Regulates Tumor Growth and Chemoresistance, A p53/lnc‐Ip53 Negative Feedback Loop Regulates Tumor Growth and Chemoresistance
Article Snippet: Protein acetylation assay was then performed as reported[ ] with modifications. .. Ten µL of affinity gel‐bound Flag, Flag‐p300 or Flag‐p300‐3 were preincubated with 1 µg of in vitro transcribed unlabeled lnc‐Ip53 or lnc‐Ip53‐AS at RT with shaking (800 r.p.m.) for 30 min, followed by adding 1 µg of purified GST‐p53, 0.5 mm acetyl coenzyme A (Ac‐CoA; A2056; Sigma‐Aldrich, St Louis, MO, USA), 2 × HAT buffer and DEPC‐H2O to make up a final volume of 30 µL reaction mixture containing 1 × HAT buffer, then incubated at 30 °C with shaking (800 r.p.m.) for 90 min before immunoblotting. .. Cell Cycle AnalysisCells were stained with propidium iodide (PI), then analyzed by fluorescence‐activated cell sorting (FACS; Gallios, Beckman Coulter, Miami, FL, USA).

Incubation:

Article Title: Contractile activity attenuates autophagy suppression and reverses mitochondrial defects in skeletal muscle cells
Article Snippet: Whole cell and mitochondrial fraction extracts (20–40 μg of protein) were separated using 8–15% SDS-PAGE and then transferred to nitrocellulose membranes. .. Membranes were blocked in 5% skim milk in TBST buffer (100 mM TRIS, 100 mM NaCl, 0.1% Tween 20 [Sigma, P1379]) for 1 h, and then incubated overnight at 4°C with primary antibodies directed against COX4I1 (1:1000; Abcam, ab14744), SOD2 (1:000; Upstate Biotechnology, 06–984), BNIP3L/NIX (1:500; Abcam, ab109414), LC3B (1:1000; Cell Signaling Technology, 2775), SQSTM1/p62 (1:40,0000; Sigma-Aldrich, P0067), PGM5/ACICULIN (1:500; in house), VDAC (1:5000; Abcam, ab1473), PPARGC1A/PGC1α (1:1000; Millipore Corporation, ab3243), p-RPS6KB (1:1000, Cell Signaling Technology, 9202), pan-LAMP2 (abcam, ab13524; 1:1000), CTSD (1:1000; Santa Cruz Biotechnology, SC6486), Tubulin (1:90,000, Calbiochem, CP06), ACTB (1:1000, Santa Cruz Biotechnology, 47,778) and TFEB (1:1000; MyBioSource, mbs120432). ..

Article Title: A p53/lnc‐Ip53 Negative Feedback Loop Regulates Tumor Growth and Chemoresistance, A p53/lnc‐Ip53 Negative Feedback Loop Regulates Tumor Growth and Chemoresistance
Article Snippet: Protein acetylation assay was then performed as reported[ ] with modifications. .. Ten µL of affinity gel‐bound Flag, Flag‐p300 or Flag‐p300‐3 were preincubated with 1 µg of in vitro transcribed unlabeled lnc‐Ip53 or lnc‐Ip53‐AS at RT with shaking (800 r.p.m.) for 30 min, followed by adding 1 µg of purified GST‐p53, 0.5 mm acetyl coenzyme A (Ac‐CoA; A2056; Sigma‐Aldrich, St Louis, MO, USA), 2 × HAT buffer and DEPC‐H2O to make up a final volume of 30 µL reaction mixture containing 1 × HAT buffer, then incubated at 30 °C with shaking (800 r.p.m.) for 90 min before immunoblotting. .. Cell Cycle AnalysisCells were stained with propidium iodide (PI), then analyzed by fluorescence‐activated cell sorting (FACS; Gallios, Beckman Coulter, Miami, FL, USA).

HAT Assay:

Article Title: A p53/lnc‐Ip53 Negative Feedback Loop Regulates Tumor Growth and Chemoresistance, A p53/lnc‐Ip53 Negative Feedback Loop Regulates Tumor Growth and Chemoresistance
Article Snippet: Protein acetylation assay was then performed as reported[ ] with modifications. .. Ten µL of affinity gel‐bound Flag, Flag‐p300 or Flag‐p300‐3 were preincubated with 1 µg of in vitro transcribed unlabeled lnc‐Ip53 or lnc‐Ip53‐AS at RT with shaking (800 r.p.m.) for 30 min, followed by adding 1 µg of purified GST‐p53, 0.5 mm acetyl coenzyme A (Ac‐CoA; A2056; Sigma‐Aldrich, St Louis, MO, USA), 2 × HAT buffer and DEPC‐H2O to make up a final volume of 30 µL reaction mixture containing 1 × HAT buffer, then incubated at 30 °C with shaking (800 r.p.m.) for 90 min before immunoblotting. .. Cell Cycle AnalysisCells were stained with propidium iodide (PI), then analyzed by fluorescence‐activated cell sorting (FACS; Gallios, Beckman Coulter, Miami, FL, USA).

Recombinant:

Article Title: S-nitrosylated SHP-2 contributes to NMDA receptor-mediated excitotoxicity in acute ischemic stroke
Article Snippet: For the biotin-switch assay, cortical hemispheres ( n = 3) were harvested from the control or ischemic side of the brain immediately after reperfusion. .. In vitro phosphatase activity of recombinant Shp-2 with pNPP (Sigma) as a substrate was measured in a 30-μL reaction mixture containing 20 mM pNPP, 50 mM sodium citrate (pH 4.8), and 1 μM recombinant SHP-2 ( ). .. After 30 min incubation at 37 °C, the reaction was terminated by addition of 1 M NaOH (270 μL).

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    Millipore egta am
    BMTs activate AMPK through CaMKK and not through intracellular Ca 2+ changes. Serum-starved L6 myotubes were treated with either vehicle (V) alone, <t>BMT-17</t> at 0.1 µM, 1 µM or 10 µM for 30 min before lysis; 10 µg of lysate was then analysed by Western blot ( A ) and quantified by densitometry ( B ). A representative blot is shown. Serum-starved L6 myotubes were pre-treated with either 150 µM <t>EGTA-AM</t> (+) or diluent (-) for 15 min before then being treated with 10 µM BMT-17 or vehicle for a further 30 min before lysis. AMPK was then isolated from 50 µg lysate by pan-AMPKβ immunoprecipitation and assessed by AMPK in vitro kinase assay ( C ). Data are means relative to untreated vehicle control (V) ± SEM from 5 independent experiments. *p
    Egta Am, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/egta am/product/Millipore
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    egta am - by Bioz Stars, 2021-03
    97/100 stars
      Buy from Supplier

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    BMTs activate AMPK through CaMKK and not through intracellular Ca 2+ changes. Serum-starved L6 myotubes were treated with either vehicle (V) alone, BMT-17 at 0.1 µM, 1 µM or 10 µM for 30 min before lysis; 10 µg of lysate was then analysed by Western blot ( A ) and quantified by densitometry ( B ). A representative blot is shown. Serum-starved L6 myotubes were pre-treated with either 150 µM EGTA-AM (+) or diluent (-) for 15 min before then being treated with 10 µM BMT-17 or vehicle for a further 30 min before lysis. AMPK was then isolated from 50 µg lysate by pan-AMPKβ immunoprecipitation and assessed by AMPK in vitro kinase assay ( C ). Data are means relative to untreated vehicle control (V) ± SEM from 5 independent experiments. *p

    Journal: PLoS ONE

    Article Title: Activation of AMPK by Bitter Melon Triterpenoids Involves CaMKK?

    doi: 10.1371/journal.pone.0062309

    Figure Lengend Snippet: BMTs activate AMPK through CaMKK and not through intracellular Ca 2+ changes. Serum-starved L6 myotubes were treated with either vehicle (V) alone, BMT-17 at 0.1 µM, 1 µM or 10 µM for 30 min before lysis; 10 µg of lysate was then analysed by Western blot ( A ) and quantified by densitometry ( B ). A representative blot is shown. Serum-starved L6 myotubes were pre-treated with either 150 µM EGTA-AM (+) or diluent (-) for 15 min before then being treated with 10 µM BMT-17 or vehicle for a further 30 min before lysis. AMPK was then isolated from 50 µg lysate by pan-AMPKβ immunoprecipitation and assessed by AMPK in vitro kinase assay ( C ). Data are means relative to untreated vehicle control (V) ± SEM from 5 independent experiments. *p

    Article Snippet: The inability of EGTA-AM to block the BMT-dependent AMPK activation demonstrated that the BMT-dependent AMPK activation was independent of intracellular calcium levels.

    Techniques: Lysis, Western Blot, Isolation, Immunoprecipitation, In Vitro, Kinase Assay

    Dynamin I dephosphorylation is arrested by BAPTA ‐ AM and EGTA ‐ AM . Cultures were preincubated with BAPTA ‐ AM , EGTA ‐ AM (both 100 μM) or DMSO (vehicle control) for 30 min. They were then either stimulated with a train of 800 action potentials (80 Hz) or left to rest for 10 s. Cultures were then immediately lysed in SDS sample buffer. (a) Representative immunoblots of cerebellar granule neuron lysates probed with either dynamin I phospho‐Ser774 ( PD ynI) or synaptophysin (Syp) antibodies. (b) Quantification of PD ynI levels after normalisation against Syp ( PD ynI/Syp) ± SEM . Blue bars represent Ctrl, red bars BAPTA ‐ AM and purple bars EGTA ‐ AM , *** p

    Journal: Journal of Neurochemistry

    Article Title: Synaptic vesicle exocytosis and increased cytosolic calcium are both necessary but not sufficient for activity‐dependent bulk endocytosis

    doi: 10.1111/jnc.13132

    Figure Lengend Snippet: Dynamin I dephosphorylation is arrested by BAPTA ‐ AM and EGTA ‐ AM . Cultures were preincubated with BAPTA ‐ AM , EGTA ‐ AM (both 100 μM) or DMSO (vehicle control) for 30 min. They were then either stimulated with a train of 800 action potentials (80 Hz) or left to rest for 10 s. Cultures were then immediately lysed in SDS sample buffer. (a) Representative immunoblots of cerebellar granule neuron lysates probed with either dynamin I phospho‐Ser774 ( PD ynI) or synaptophysin (Syp) antibodies. (b) Quantification of PD ynI levels after normalisation against Syp ( PD ynI/Syp) ± SEM . Blue bars represent Ctrl, red bars BAPTA ‐ AM and purple bars EGTA ‐ AM , *** p

    Article Snippet: The inhibition of dynamin I dephosphorylation in the presence of EGTA‐AM is consistent with this event being central in the triggering of this endocytosis mode.

    Techniques: De-Phosphorylation Assay, Western Blot

    Activity‐dependent bulk endocytosis ( ADBE ) is abolished by both BAPTA ‐ AM and EGTA ‐ AM . Cultures were preincubated with BAPTA ‐ AM , EGTA ‐ AM (both 100 μM) or DMSO (vehicle control) for 30 min before loading with tetramethylrhodamine‐dextran ( TMR )‐dextran (50 μM) during stimulation with a train of 800 action potentials (80 Hz). (a) Quantification of the number of evoked TMR ‐dextran puncta per field of view in cultures treated with either DMSO (Ctrl, blue bars) BAPTA ‐ AM (red bars) or EGTA ‐ AM (purple bars) ± SEM . *** p

    Journal: Journal of Neurochemistry

    Article Title: Synaptic vesicle exocytosis and increased cytosolic calcium are both necessary but not sufficient for activity‐dependent bulk endocytosis

    doi: 10.1111/jnc.13132

    Figure Lengend Snippet: Activity‐dependent bulk endocytosis ( ADBE ) is abolished by both BAPTA ‐ AM and EGTA ‐ AM . Cultures were preincubated with BAPTA ‐ AM , EGTA ‐ AM (both 100 μM) or DMSO (vehicle control) for 30 min before loading with tetramethylrhodamine‐dextran ( TMR )‐dextran (50 μM) during stimulation with a train of 800 action potentials (80 Hz). (a) Quantification of the number of evoked TMR ‐dextran puncta per field of view in cultures treated with either DMSO (Ctrl, blue bars) BAPTA ‐ AM (red bars) or EGTA ‐ AM (purple bars) ± SEM . *** p

    Article Snippet: EGTA‐AM, BAPTA‐AM (both 100 μM) or an equal volume of vehicle (DMSO) was incubated with cultures after the S2 load for 30 min before the S2 unloading step.

    Techniques: Activity Assay

    SV exocytosis is arrested by BAPTA ‐ AM but not by EGTA ‐ AM . (a) Schematic of the experimental design. Cultures were loaded with 10 μM FM 1‐43 during a train of 800 action potentials (80 Hz). After a 10 min rest period FM 1‐43 was unloaded with a second 800 action potential stimulus (S1). After a 10 min rest period, the same loading/unloading protocol was performed again, with the exception being that cultures were incubated with either BAPTA ‐ AM , EGTA ‐ AM (both 100 μM) or dimethylsulfoxide ( DMSO ) (Ctrl) for 30 min directly after S2 loading. (b) Representative traces showing the S2 unloading profiles of Ctrl (blue circles), BAPTA ‐ AM (red circles) and EGTA ‐ AM (purple circles) ‐treated cultures. (c) Quantification of the extent of FM 1‐43 unloading at both S1 (ΔS1) and S2 (ΔS2) presented as the ratio ΔS2/ΔS1 ± SEM . *** p

    Journal: Journal of Neurochemistry

    Article Title: Synaptic vesicle exocytosis and increased cytosolic calcium are both necessary but not sufficient for activity‐dependent bulk endocytosis

    doi: 10.1111/jnc.13132

    Figure Lengend Snippet: SV exocytosis is arrested by BAPTA ‐ AM but not by EGTA ‐ AM . (a) Schematic of the experimental design. Cultures were loaded with 10 μM FM 1‐43 during a train of 800 action potentials (80 Hz). After a 10 min rest period FM 1‐43 was unloaded with a second 800 action potential stimulus (S1). After a 10 min rest period, the same loading/unloading protocol was performed again, with the exception being that cultures were incubated with either BAPTA ‐ AM , EGTA ‐ AM (both 100 μM) or dimethylsulfoxide ( DMSO ) (Ctrl) for 30 min directly after S2 loading. (b) Representative traces showing the S2 unloading profiles of Ctrl (blue circles), BAPTA ‐ AM (red circles) and EGTA ‐ AM (purple circles) ‐treated cultures. (c) Quantification of the extent of FM 1‐43 unloading at both S1 (ΔS1) and S2 (ΔS2) presented as the ratio ΔS2/ΔS1 ± SEM . *** p

    Article Snippet: EGTA‐AM, BAPTA‐AM (both 100 μM) or an equal volume of vehicle (DMSO) was incubated with cultures after the S2 load for 30 min before the S2 unloading step.

    Techniques: Incubation