ethylene glycol tetraacetic acid egta  (Thermo Fisher)


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    Name:
    Ethylene Glycol
    Description:
    Thermo Scientific Pierce Ethylene Glycol Solution provides exceptional antifreeze protection and storage stability for antibody enzyme conjugates because it is purified to remove impurities commonly found traditional glycerol stocks Features of Ethylene Glycol • Specially purified to remove impurities such as aldehydes peroxides iron and UV absorbing hydrocarbons • Suitable for enzyme storage without the worry of losing enzymatic activity • Stable for months This product is a 50 w v aqueous solution of highly purified ethylene glycol When mixed in equal volume with purified protein samples such as primary antibodies the solution stabilizes and maintains the mixture as a liquid during freezer storage 20° Ethylene glycol is a suitable alternative to glycerol for most protein storage applications In fact this preparation of ethylene glycol is devoid of various common glycerol impurities and potiential degradation products that are damaging to certain enzymes
    Catalog Number:
    29810
    Price:
    None
    Applications:
    Antibody Labeling|Antibody Production and Labeling|Build Your Own Immunoassay|Cell Analysis|ELISA|Protein Assays and Analysis|Protein Biology|Western Blotting
    Category:
    Lab Reagents and Chemicals
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    Structured Review

    Thermo Fisher ethylene glycol tetraacetic acid egta
    Thermo Scientific Pierce Ethylene Glycol Solution provides exceptional antifreeze protection and storage stability for antibody enzyme conjugates because it is purified to remove impurities commonly found traditional glycerol stocks Features of Ethylene Glycol • Specially purified to remove impurities such as aldehydes peroxides iron and UV absorbing hydrocarbons • Suitable for enzyme storage without the worry of losing enzymatic activity • Stable for months This product is a 50 w v aqueous solution of highly purified ethylene glycol When mixed in equal volume with purified protein samples such as primary antibodies the solution stabilizes and maintains the mixture as a liquid during freezer storage 20° Ethylene glycol is a suitable alternative to glycerol for most protein storage applications In fact this preparation of ethylene glycol is devoid of various common glycerol impurities and potiential degradation products that are damaging to certain enzymes
    https://www.bioz.com/result/ethylene glycol tetraacetic acid egta/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ethylene glycol tetraacetic acid egta - by Bioz Stars, 2020-07
    99/100 stars

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    Related Articles

    Isolation:

    Article Title: Regulation of inflammation and fibrosis by macrophages in lymphedema
    Article Snippet: .. VEGF-C protein expression was quantified in total protein isolated from the tail tissues using ELISA (eBiosciences) as previously described ( ). .. Briefly, skin and subcutaneous tissues were harvested and lysed using the Qiagen DNA/RNA/Protein mini kit using the manufacturer's methods (Qiagen, Valencia, CA).

    Labeling:

    Article Title: ATTACK, a novel bispecific T cell-recruiting antibody with trivalent EGFR binding and monovalent CD3 binding for cancer immunotherapy
    Article Snippet: .. Immunological synapse formation and polarized PLCγ signalling For imaging, 3T3 and 3T3-EGFR cells were fluorescently labeled with 10 µM cell tracker dye 7-amino-4-chloromethylcoumarin (CMAC; Life Technologies) for 30 minutes and incubated with purified EgA1 LiTE or EgA1 ATTACK at concentrations ranging from 50 pM to 5 nM for one hour. ..

    Article Title: Selective targeting of melanoma by PEG-masked protein-based multifunctional nanoparticles
    Article Snippet: .. Fluorescent labeling of proteins HFt-PEG and HFt-MSH-PEG solutions (2 mg/mL) were incubated with 1 mM of 5(6)-carboxytetramethylrhodamine N-succinimidyl ester (λex 552 nm, λem 575 nm; Thermo Scientific, Milan, Italy) in phosphate-buffered saline for 2 hours at pH 7.5 and room temperature, with stirring in the dark. .. Subsequently, the samples were filtered, dialyzed, and exchanged with H2 Odd and phosphate-buffered saline, using 30 kDa Amicon Ultra-15 centrifugal filter devices to remove excess reagents.

    Purification:

    Article Title: ATTACK, a novel bispecific T cell-recruiting antibody with trivalent EGFR binding and monovalent CD3 binding for cancer immunotherapy
    Article Snippet: .. Immunological synapse formation and polarized PLCγ signalling For imaging, 3T3 and 3T3-EGFR cells were fluorescently labeled with 10 µM cell tracker dye 7-amino-4-chloromethylcoumarin (CMAC; Life Technologies) for 30 minutes and incubated with purified EgA1 LiTE or EgA1 ATTACK at concentrations ranging from 50 pM to 5 nM for one hour. ..

    Enzyme-linked Immunosorbent Assay:

    Article Title: Regulation of inflammation and fibrosis by macrophages in lymphedema
    Article Snippet: .. VEGF-C protein expression was quantified in total protein isolated from the tail tissues using ELISA (eBiosciences) as previously described ( ). .. Briefly, skin and subcutaneous tissues were harvested and lysed using the Qiagen DNA/RNA/Protein mini kit using the manufacturer's methods (Qiagen, Valencia, CA).

    Article Title: Differential regulation of TGF-β–induced, ALK-5–mediated VEGF release by SMAD2/3 versus SMAD1/5/8 signaling in glioblastoma
    Article Snippet: .. ) were analyzed by ELISA for VEGF levels (eBioscience) and TGF-β1/2 (R & D Systems). .. The pGL3 SBE-4-Luc plasmid (B. Vogelstein) containing 4 copies of the SMAD-binding element (SBE) GTCTAGAC ).

    Article Title: Elevated VEGF Levels in Pulmonary Edema Fluid and PBMCs from Patients with Acute Hantavirus Pulmonary Syndrome
    Article Snippet: .. ELISA Assay VEGF levels in samples were blindly assessed using a quantitative sandwich ELISA performed with the Human VEGF ELISA kit (Thermo Scientific) with a sensitivity of VEGF detection > 8 pg/mL. .. Briefly, VEGF levels were determined following capture with a solid phase monoclonal antibody to VEGF165 IgG and detected using a biotin-conjugated polyclonal rabbit antibody against recombinant human VEGF165 .

    Article Title: Ghrelin Inhibits the Differentiation of T Helper 17 Cells through mTOR/STAT3 Signaling Pathway
    Article Snippet: .. ELISA Analysis Supernatant of cell culture or serum of mice was collected, ELISA was performed with mouse IL-17A quantifying kit (eBioscience, San Diego, CA) with an indicated sensitivity of below 4 pg/ml. .. Western Blot Analysis Immunoblotting was performed as described previously.

    Article Title: Brain-Targeted Proanthocyanidin Metabolites for Alzheimer\u2019s Disease Treatment
    Article Snippet: .. Specifically, soluble amyloid peptide was extracted in PBS and centrifuged at 78,500 × g for 1 h at 4 °C, and the supernatant was quantified by ELISA to specifically detect aggregated Aβ (Invitrogen). ..

    Sandwich ELISA:

    Article Title: Elevated VEGF Levels in Pulmonary Edema Fluid and PBMCs from Patients with Acute Hantavirus Pulmonary Syndrome
    Article Snippet: .. ELISA Assay VEGF levels in samples were blindly assessed using a quantitative sandwich ELISA performed with the Human VEGF ELISA kit (Thermo Scientific) with a sensitivity of VEGF detection > 8 pg/mL. .. Briefly, VEGF levels were determined following capture with a solid phase monoclonal antibody to VEGF165 IgG and detected using a biotin-conjugated polyclonal rabbit antibody against recombinant human VEGF165 .

    Incubation:

    Article Title: ATTACK, a novel bispecific T cell-recruiting antibody with trivalent EGFR binding and monovalent CD3 binding for cancer immunotherapy
    Article Snippet: .. Immunological synapse formation and polarized PLCγ signalling For imaging, 3T3 and 3T3-EGFR cells were fluorescently labeled with 10 µM cell tracker dye 7-amino-4-chloromethylcoumarin (CMAC; Life Technologies) for 30 minutes and incubated with purified EgA1 LiTE or EgA1 ATTACK at concentrations ranging from 50 pM to 5 nM for one hour. ..

    Article Title: Selective targeting of melanoma by PEG-masked protein-based multifunctional nanoparticles
    Article Snippet: .. Fluorescent labeling of proteins HFt-PEG and HFt-MSH-PEG solutions (2 mg/mL) were incubated with 1 mM of 5(6)-carboxytetramethylrhodamine N-succinimidyl ester (λex 552 nm, λem 575 nm; Thermo Scientific, Milan, Italy) in phosphate-buffered saline for 2 hours at pH 7.5 and room temperature, with stirring in the dark. .. Subsequently, the samples were filtered, dialyzed, and exchanged with H2 Odd and phosphate-buffered saline, using 30 kDa Amicon Ultra-15 centrifugal filter devices to remove excess reagents.

    Cell Culture:

    Article Title: Ghrelin Inhibits the Differentiation of T Helper 17 Cells through mTOR/STAT3 Signaling Pathway
    Article Snippet: .. ELISA Analysis Supernatant of cell culture or serum of mice was collected, ELISA was performed with mouse IL-17A quantifying kit (eBioscience, San Diego, CA) with an indicated sensitivity of below 4 pg/ml. .. Western Blot Analysis Immunoblotting was performed as described previously.

    Expressing:

    Article Title: Regulation of inflammation and fibrosis by macrophages in lymphedema
    Article Snippet: .. VEGF-C protein expression was quantified in total protein isolated from the tail tissues using ELISA (eBiosciences) as previously described ( ). .. Briefly, skin and subcutaneous tissues were harvested and lysed using the Qiagen DNA/RNA/Protein mini kit using the manufacturer's methods (Qiagen, Valencia, CA).

    Mouse Assay:

    Article Title: Ghrelin Inhibits the Differentiation of T Helper 17 Cells through mTOR/STAT3 Signaling Pathway
    Article Snippet: .. ELISA Analysis Supernatant of cell culture or serum of mice was collected, ELISA was performed with mouse IL-17A quantifying kit (eBioscience, San Diego, CA) with an indicated sensitivity of below 4 pg/ml. .. Western Blot Analysis Immunoblotting was performed as described previously.

    Imaging:

    Article Title: ATTACK, a novel bispecific T cell-recruiting antibody with trivalent EGFR binding and monovalent CD3 binding for cancer immunotherapy
    Article Snippet: .. Immunological synapse formation and polarized PLCγ signalling For imaging, 3T3 and 3T3-EGFR cells were fluorescently labeled with 10 µM cell tracker dye 7-amino-4-chloromethylcoumarin (CMAC; Life Technologies) for 30 minutes and incubated with purified EgA1 LiTE or EgA1 ATTACK at concentrations ranging from 50 pM to 5 nM for one hour. ..

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    Thermo Fisher caspase lysis buffer
    Statin-induced apoptosis and its lipid dependency in NSCLC cells. ( A ) Evaluation of <t>caspase</t> 3 activation in cells treated with up to 100 µM of either pitavastatin or fluvastatin for 48 h. Mevalonic acid (Mev, 1 mM) was used as rescue control counteracting statin-mediated intracellular depletion of sterol precursors. Mev and statin co-treated cells showed caspase 3 activity indistinguishable from untreated cells. One-way ANOVA, α = 0.05; Calu6 (pitavastatin): F(9,22) = 4.805, p = 0.0013; Calu6 (fluvastatin): F(9,16) = 2.207, p = 0.0801; H1993 (pitavastatin): F(9,26) = 1.337, p = 0.2664; H1993 (fluvastatin): F(9,21) = 2.794, p = 0.0252; A549 (pitavastatin): F(9,30) = 4.152, p = 0.0015; A549 (fluvastatin): F(9,18) = 3.411, p = 0.0128. Data are given as arithmetic mean of fold change relative to untreated control (CTL) ± SD from three independent experiments. Asterisks denote statistical significance as determined via Dunnett’s multiple comparison test compared to untreated control (CTL): *p
    Caspase Lysis Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 3 article reviews
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    92
    Thermo Fisher csf xb buffer
    Characterization of pSer335 <t>XErp1</t> antibody <t>CSF</t> extract was treated with Myc‐XErp1 IVT carrying the indicated combinations of the mutations DSG − (S33N S38N), DSA − (S284N S288N), ZBR − (C583A) and CaMKII − (T195A). Calcium was added, samples were taken at the indicated time points and as indicated treated with λ‐phosphatase. Samples were immunoblotted for the Myc‐tag and cyclin B2. The cyclin B2 membrane was stripped and reprobed for α‐tubulin. Asterisk indicates unspecific bands. Several lanes were removed at the dashed line. CSF extract was treated with Myc‐XErp1 CaMKII − ZBR − (T195A C583A) IVT at the indicated dilutions. An empty IVT not expressing XErp1 and an untreated condition were used as controls. Samples were taken, treated as indicated with λ‐phosphatase and immunoblotted for XErp1, pSer335 XErp1, and α‐tubulin. The XErp1 membrane was stripped and reprobed for the Myc‐tag. Asterisks indicate unspecific bands. CSF extract was treated with Myc‐XErp1 CaMKII − ZBR − (T195A C583A) IVT that was either wild‐type or mutated to alanine at Ser335. An empty IVT reaction not expressing Myc‐XErp1 served as control. Samples were taken, treated as indicated with λ‐phosphatase and immunoblotted for XErp1, the Myc‐tag, pSer335 XErp1, and α‐tubulin. Asterisks indicate unspecific bands. Source data are available online for this figure.
    Csf Xb Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    thermo fisher egta am
    The suppression of Ca 2+ puffs during global signals does not result because of elevated cytosolic [Ca 2+ ]. ( A,B ) IP 3 -evoked Ca 2+ puffs are not suppressed by prior photorelease of Ca 2+ . ( A ) Traces depict fluorescence ratios (black; ΔF/F 0 ) from WT HEK cells and corresponding SD signals (grey; in arbitrary units, A.U.). Records, from left to right, show responses from individual cells loaded with <t>NP-EGTA</t> (caged Ca 2+ ) that were unstimulated or exposed to increasing UV flash durations (marked by arrows) to photorelease progressively increasing amounts of free Ca 2+ before challenging cells with CCH (100 µM) locally delivered by a puffer pipette when indicated by the bars. Traces are blanked out during the artifact caused by the photolysis flash. ( B ) Data points from traces like those in A show the integral under SD trace (a measure of puff activity) as a ratio of the change in global Ca 2+ signal (ΔF/F 0 ) evoked by CCH. The data are binned in terms of the jump in <t>Cal520</t> fluorescence (ΔF/F 0 ) evoked by photolysis of caged Ca 2+ . Open symbols are from individual cells, and filled symbols are means for each group (respective n numbers for different bins; 20, 4, 4, 6, 8). Data are normalized with respect to the mean ratio without prior photorelease of Ca 2+ . There was no significant difference between control CCH responses and CCH responses following Ca 2+ jumps (evaluated by Student T-test; p values between 0.17 and 0.66 for the different binned groupings). ( C,D ) Termination of puff activity is unaffected when global cytosolic Ca 2+ signals are attenuated by buffering with EGTA. ( C ) Traces showing the Cal520 fluorescence ratio (ΔF/F 0 ; smooth trace) and SD signal (noisy trace) in response to photoreleased i-IP 3 in a representative WT HEK cell that was incubated with 15 µM EGTA/AM to buffer cytosolic Ca 2+ and attenuate the amplitude of the global Ca 2+ signal. ( D ) Scatter plots show measurements of the SD signal at intervals during the rising phase of global Ca 2+ responses against the magnitude of the global Ca 2+ elevation (ΔF/F 0 ) at that time. Measurements were binned at intervals of (0.1 ΔF/F 0 ) and SD data are normalized to a peak value of 1. Solid circles show mean data from 14 EGTA-loaded cells. For comparison, open circles present data reproduced from Figure 3C showing measurements from 11 control cells that gave fast rising responses to photoreleased i-IP 3 .
    Egta Am, supplied by thermo fisher, used in various techniques. Bioz Stars score: 92/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Statin-induced apoptosis and its lipid dependency in NSCLC cells. ( A ) Evaluation of caspase 3 activation in cells treated with up to 100 µM of either pitavastatin or fluvastatin for 48 h. Mevalonic acid (Mev, 1 mM) was used as rescue control counteracting statin-mediated intracellular depletion of sterol precursors. Mev and statin co-treated cells showed caspase 3 activity indistinguishable from untreated cells. One-way ANOVA, α = 0.05; Calu6 (pitavastatin): F(9,22) = 4.805, p = 0.0013; Calu6 (fluvastatin): F(9,16) = 2.207, p = 0.0801; H1993 (pitavastatin): F(9,26) = 1.337, p = 0.2664; H1993 (fluvastatin): F(9,21) = 2.794, p = 0.0252; A549 (pitavastatin): F(9,30) = 4.152, p = 0.0015; A549 (fluvastatin): F(9,18) = 3.411, p = 0.0128. Data are given as arithmetic mean of fold change relative to untreated control (CTL) ± SD from three independent experiments. Asterisks denote statistical significance as determined via Dunnett’s multiple comparison test compared to untreated control (CTL): *p

    Journal: Scientific Reports

    Article Title: Delineation of cell death mechanisms induced by synergistic effects of statins and erlotinib in non-small cell lung cancer cell (NSCLC) lines

    doi: 10.1038/s41598-020-57707-2

    Figure Lengend Snippet: Statin-induced apoptosis and its lipid dependency in NSCLC cells. ( A ) Evaluation of caspase 3 activation in cells treated with up to 100 µM of either pitavastatin or fluvastatin for 48 h. Mevalonic acid (Mev, 1 mM) was used as rescue control counteracting statin-mediated intracellular depletion of sterol precursors. Mev and statin co-treated cells showed caspase 3 activity indistinguishable from untreated cells. One-way ANOVA, α = 0.05; Calu6 (pitavastatin): F(9,22) = 4.805, p = 0.0013; Calu6 (fluvastatin): F(9,16) = 2.207, p = 0.0801; H1993 (pitavastatin): F(9,26) = 1.337, p = 0.2664; H1993 (fluvastatin): F(9,21) = 2.794, p = 0.0252; A549 (pitavastatin): F(9,30) = 4.152, p = 0.0015; A549 (fluvastatin): F(9,18) = 3.411, p = 0.0128. Data are given as arithmetic mean of fold change relative to untreated control (CTL) ± SD from three independent experiments. Asterisks denote statistical significance as determined via Dunnett’s multiple comparison test compared to untreated control (CTL): *p

    Article Snippet: Caspase lysis buffer (25 mM HEPES, 5 mM EDTA, 1 mM EGTA, 5 mM MgCl2 ) was used as a background control.

    Techniques: Activation Assay, Activity Assay

    Characterization of pSer335 XErp1 antibody CSF extract was treated with Myc‐XErp1 IVT carrying the indicated combinations of the mutations DSG − (S33N S38N), DSA − (S284N S288N), ZBR − (C583A) and CaMKII − (T195A). Calcium was added, samples were taken at the indicated time points and as indicated treated with λ‐phosphatase. Samples were immunoblotted for the Myc‐tag and cyclin B2. The cyclin B2 membrane was stripped and reprobed for α‐tubulin. Asterisk indicates unspecific bands. Several lanes were removed at the dashed line. CSF extract was treated with Myc‐XErp1 CaMKII − ZBR − (T195A C583A) IVT at the indicated dilutions. An empty IVT not expressing XErp1 and an untreated condition were used as controls. Samples were taken, treated as indicated with λ‐phosphatase and immunoblotted for XErp1, pSer335 XErp1, and α‐tubulin. The XErp1 membrane was stripped and reprobed for the Myc‐tag. Asterisks indicate unspecific bands. CSF extract was treated with Myc‐XErp1 CaMKII − ZBR − (T195A C583A) IVT that was either wild‐type or mutated to alanine at Ser335. An empty IVT reaction not expressing Myc‐XErp1 served as control. Samples were taken, treated as indicated with λ‐phosphatase and immunoblotted for XErp1, the Myc‐tag, pSer335 XErp1, and α‐tubulin. Asterisks indicate unspecific bands. Source data are available online for this figure.

    Journal: EMBO Reports

    Article Title: Calcineurin promotes APC/C activation at meiotic exit by acting on both XErp1 and Cdc20

    doi: 10.15252/embr.201846433

    Figure Lengend Snippet: Characterization of pSer335 XErp1 antibody CSF extract was treated with Myc‐XErp1 IVT carrying the indicated combinations of the mutations DSG − (S33N S38N), DSA − (S284N S288N), ZBR − (C583A) and CaMKII − (T195A). Calcium was added, samples were taken at the indicated time points and as indicated treated with λ‐phosphatase. Samples were immunoblotted for the Myc‐tag and cyclin B2. The cyclin B2 membrane was stripped and reprobed for α‐tubulin. Asterisk indicates unspecific bands. Several lanes were removed at the dashed line. CSF extract was treated with Myc‐XErp1 CaMKII − ZBR − (T195A C583A) IVT at the indicated dilutions. An empty IVT not expressing XErp1 and an untreated condition were used as controls. Samples were taken, treated as indicated with λ‐phosphatase and immunoblotted for XErp1, pSer335 XErp1, and α‐tubulin. The XErp1 membrane was stripped and reprobed for the Myc‐tag. Asterisks indicate unspecific bands. CSF extract was treated with Myc‐XErp1 CaMKII − ZBR − (T195A C583A) IVT that was either wild‐type or mutated to alanine at Ser335. An empty IVT reaction not expressing Myc‐XErp1 served as control. Samples were taken, treated as indicated with λ‐phosphatase and immunoblotted for XErp1, the Myc‐tag, pSer335 XErp1, and α‐tubulin. Asterisks indicate unspecific bands. Source data are available online for this figure.

    Article Snippet: XErp1: 2 μl Myc‐XErp1 IVT were diluted in 8 μl CSF‐XB Buffer (10 mM HEPES; 100 mM KCl; 2 mM MgCl2 ; 0.1 mM CaCl2 ; 50 mM sucrose; 5 mM EGTA; pH = 7.7) and added to 0.5 μg α‐Myc antibodies coupled to Dynabeads® Protein G (Thermo Fisher).

    Techniques: Expressing

    The suppression of Ca 2+ puffs during global signals does not result because of elevated cytosolic [Ca 2+ ]. ( A,B ) IP 3 -evoked Ca 2+ puffs are not suppressed by prior photorelease of Ca 2+ . ( A ) Traces depict fluorescence ratios (black; ΔF/F 0 ) from WT HEK cells and corresponding SD signals (grey; in arbitrary units, A.U.). Records, from left to right, show responses from individual cells loaded with NP-EGTA (caged Ca 2+ ) that were unstimulated or exposed to increasing UV flash durations (marked by arrows) to photorelease progressively increasing amounts of free Ca 2+ before challenging cells with CCH (100 µM) locally delivered by a puffer pipette when indicated by the bars. Traces are blanked out during the artifact caused by the photolysis flash. ( B ) Data points from traces like those in A show the integral under SD trace (a measure of puff activity) as a ratio of the change in global Ca 2+ signal (ΔF/F 0 ) evoked by CCH. The data are binned in terms of the jump in Cal520 fluorescence (ΔF/F 0 ) evoked by photolysis of caged Ca 2+ . Open symbols are from individual cells, and filled symbols are means for each group (respective n numbers for different bins; 20, 4, 4, 6, 8). Data are normalized with respect to the mean ratio without prior photorelease of Ca 2+ . There was no significant difference between control CCH responses and CCH responses following Ca 2+ jumps (evaluated by Student T-test; p values between 0.17 and 0.66 for the different binned groupings). ( C,D ) Termination of puff activity is unaffected when global cytosolic Ca 2+ signals are attenuated by buffering with EGTA. ( C ) Traces showing the Cal520 fluorescence ratio (ΔF/F 0 ; smooth trace) and SD signal (noisy trace) in response to photoreleased i-IP 3 in a representative WT HEK cell that was incubated with 15 µM EGTA/AM to buffer cytosolic Ca 2+ and attenuate the amplitude of the global Ca 2+ signal. ( D ) Scatter plots show measurements of the SD signal at intervals during the rising phase of global Ca 2+ responses against the magnitude of the global Ca 2+ elevation (ΔF/F 0 ) at that time. Measurements were binned at intervals of (0.1 ΔF/F 0 ) and SD data are normalized to a peak value of 1. Solid circles show mean data from 14 EGTA-loaded cells. For comparison, open circles present data reproduced from Figure 3C showing measurements from 11 control cells that gave fast rising responses to photoreleased i-IP 3 .

    Journal: eLife

    Article Title: IP3 mediated global Ca2+ signals arise through two temporally and spatially distinct modes of Ca2+ release

    doi: 10.7554/eLife.55008

    Figure Lengend Snippet: The suppression of Ca 2+ puffs during global signals does not result because of elevated cytosolic [Ca 2+ ]. ( A,B ) IP 3 -evoked Ca 2+ puffs are not suppressed by prior photorelease of Ca 2+ . ( A ) Traces depict fluorescence ratios (black; ΔF/F 0 ) from WT HEK cells and corresponding SD signals (grey; in arbitrary units, A.U.). Records, from left to right, show responses from individual cells loaded with NP-EGTA (caged Ca 2+ ) that were unstimulated or exposed to increasing UV flash durations (marked by arrows) to photorelease progressively increasing amounts of free Ca 2+ before challenging cells with CCH (100 µM) locally delivered by a puffer pipette when indicated by the bars. Traces are blanked out during the artifact caused by the photolysis flash. ( B ) Data points from traces like those in A show the integral under SD trace (a measure of puff activity) as a ratio of the change in global Ca 2+ signal (ΔF/F 0 ) evoked by CCH. The data are binned in terms of the jump in Cal520 fluorescence (ΔF/F 0 ) evoked by photolysis of caged Ca 2+ . Open symbols are from individual cells, and filled symbols are means for each group (respective n numbers for different bins; 20, 4, 4, 6, 8). Data are normalized with respect to the mean ratio without prior photorelease of Ca 2+ . There was no significant difference between control CCH responses and CCH responses following Ca 2+ jumps (evaluated by Student T-test; p values between 0.17 and 0.66 for the different binned groupings). ( C,D ) Termination of puff activity is unaffected when global cytosolic Ca 2+ signals are attenuated by buffering with EGTA. ( C ) Traces showing the Cal520 fluorescence ratio (ΔF/F 0 ; smooth trace) and SD signal (noisy trace) in response to photoreleased i-IP 3 in a representative WT HEK cell that was incubated with 15 µM EGTA/AM to buffer cytosolic Ca 2+ and attenuate the amplitude of the global Ca 2+ signal. ( D ) Scatter plots show measurements of the SD signal at intervals during the rising phase of global Ca 2+ responses against the magnitude of the global Ca 2+ elevation (ΔF/F 0 ) at that time. Measurements were binned at intervals of (0.1 ΔF/F 0 ) and SD data are normalized to a peak value of 1. Solid circles show mean data from 14 EGTA-loaded cells. For comparison, open circles present data reproduced from Figure 3C showing measurements from 11 control cells that gave fast rising responses to photoreleased i-IP 3 .

    Article Snippet: Cal520/AM, ci-IP3/PM, and NP-EGTA/AM, EGTA/AM, and fluo8L/AM were all solubilized with DMSO/20% pluronic F127 (Thermo Fisher Scientific #P3000MP).

    Techniques: Fluorescence, Transferring, Activity Assay, Incubation

    Cytosolic Ca 2+ signal is essential in mediating the autophagy defect induced by FAD-PSEN2 expression. (a) Representative images and quantification of LC3 dots, revealed by immuno-staining, in SH-SY5Y cells incubated, or not (control), with BAPTA-AM or EGTA-AM (2 μM, 1 h) and treated, or not (not-treated, NT) with BafA (100 nM). Mean ± SEM, n = 26–51 cells, from 3 independent experiments. *p

    Journal: Autophagy

    Article Title: PSEN2 (presenilin 2) mutants linked to familial Alzheimer disease impair autophagy by altering Ca2+ homeostasis

    doi: 10.1080/15548627.2019.1596489

    Figure Lengend Snippet: Cytosolic Ca 2+ signal is essential in mediating the autophagy defect induced by FAD-PSEN2 expression. (a) Representative images and quantification of LC3 dots, revealed by immuno-staining, in SH-SY5Y cells incubated, or not (control), with BAPTA-AM or EGTA-AM (2 μM, 1 h) and treated, or not (not-treated, NT) with BafA (100 nM). Mean ± SEM, n = 26–51 cells, from 3 independent experiments. *p

    Article Snippet: EGTA-AM or BAPTA-AM (Thermo Fisher Scientific, E1219 and B6769, respectively) treatments were performed by incubating cells for the indicated time with 2 or 5 μM of each compound, while over night (on) EGTA incubation was performed by adding 2 mM EGTA (Thermo Fisher Scientific, E3889) (pH 7.4) and 2 mM MgCl2 directly to the cell medium.

    Techniques: Expressing, Immunostaining, Incubation