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  • 85
    Millipore fura pe3
    Effect of La 3+ on HPV in IPA A and B , tension and <t>fura</t> <t>PE-3</t> F 340/380 fluorescence ratio, respectively, measured simultaneously in a single IPA under control conditions and in the presence of 1 and 100 μM La 3+ . C and D , mean results for five IPAs: control, ○; 1 μM La 3+ , •; and 100 μM La 3+ , ▪. * P
    Fura Pe3, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Immuno-Biological Laboratories Co Ltd aβ pe3
    Effect of La 3+ on HPV in IPA A and B , tension and <t>fura</t> <t>PE-3</t> F 340/380 fluorescence ratio, respectively, measured simultaneously in a single IPA under control conditions and in the presence of 1 and 100 μM La 3+ . C and D , mean results for five IPAs: control, ○; 1 μM La 3+ , •; and 100 μM La 3+ , ▪. * P
    Aβ Pe3, supplied by Immuno-Biological Laboratories Co Ltd, used in various techniques. Bioz Stars score: 90/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Probiodrug pe3 aβ
    In a therapeutic study initiated well after the onset of cerebral <t>Aβ</t> deposition and gliosis, weekly passive immunization with <t>anti-pE3-Aβ</t> mAb07/1 in 23-monthold APPswe/PS1ΔE9 mice for 7 weeks resulted in the attenuation of pE3-Aβ and general Aβ (R1282 IR) deposition as well as fibrillar amyloid (Thioflavin S) in the hippocampus ( a , b ) and cerebellum ( c , d ) compared to PBS control mice. Immunohistochemical results ( a , c ) and Thioflavin S labeling were quantified by image analysis ( b , d ). Absolute values are provided in table 1. Scale bars, 200 µm. p values: * p > 0.05; *** p > 0.001; n.s. = nonsignificant (p = 0.11); p = 0.06 (strong trend).
    Pe3 Aβ, supplied by Probiodrug, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher fura 2 pe3
    Effect of prolonged exposure to insulin on [Ca 2+ ] i , total calcium current (I Ca 2+ ), and SERCA2 protein. (A and B) INS1E β -cells cultured on glass coverslips were loaded with <t>fura-2–PE3</t> (2 µmol/L) and Mag–fluo-4-AM (1 μM), respectively (Molecular Probes, Eugene, OR). Pluronic acid (0.01%) was added to the medium and cells were incubated for 30 min at 37°C. [Ca 2+ ] i was measured using an Olympus IX-50 inverted epifluorescence microscope (Olympus, Tokyo, Japan). Absolute [Ca 2+ ] i was determined from R of Ca 2+ -bound fura-2 (excited at 340 nm) to unbound fura-2 (excited at 380 nM). The Ca 2+ levels were determined using a standard equation for calibration. R max and R min were obtained by adding ionomycin (10 µM) or EGTA (1 mM), respectively, to fura-2–loaded β -cells at the end of each experiment. [Ca 2+ ] i was obtained using an excitation wavelength of 488 nm and an emission band-pass filter of 515/15 nm. Means ± SEM are from four independent experiments. The I Ca 2+ (Ba 2+ ) from control and insulin pretreated cells for 48 h were evoked by a 100 msec step depolarization under whole-cell voltage clamp configuration. * P
    Fura 2 Pe3, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Santa Cruz Biotechnology fura pe3
    Effect of prolonged exposure to insulin on [Ca 2+ ] i , total calcium current (I Ca 2+ ), and SERCA2 protein. (A and B) INS1E β -cells cultured on glass coverslips were loaded with <t>fura-2–PE3</t> (2 µmol/L) and Mag–fluo-4-AM (1 μM), respectively (Molecular Probes, Eugene, OR). Pluronic acid (0.01%) was added to the medium and cells were incubated for 30 min at 37°C. [Ca 2+ ] i was measured using an Olympus IX-50 inverted epifluorescence microscope (Olympus, Tokyo, Japan). Absolute [Ca 2+ ] i was determined from R of Ca 2+ -bound fura-2 (excited at 340 nm) to unbound fura-2 (excited at 380 nM). The Ca 2+ levels were determined using a standard equation for calibration. R max and R min were obtained by adding ionomycin (10 µM) or EGTA (1 mM), respectively, to fura-2–loaded β -cells at the end of each experiment. [Ca 2+ ] i was obtained using an excitation wavelength of 488 nm and an emission band-pass filter of 515/15 nm. Means ± SEM are from four independent experiments. The I Ca 2+ (Ba 2+ ) from control and insulin pretreated cells for 48 h were evoked by a 100 msec step depolarization under whole-cell voltage clamp configuration. * P
    Fura Pe3, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Millipore fura pe3 am
    Lysophosphatidic acid (LPA)-induced Ca 2+ entry requires sufficient levels of STIM1 and functional Orai1. ( a – c ) Relative efficiency of small interfering RNA (siRNA)-mediated STIM1 silencing compared to scrambled (scr) control by quantitative PCR and western blotting. ( e ) <t>Fura-PE3-based</t> Ca 2+ imaging showed impaired LPA-induced Ca 2+ entry in STIM1-knockdown cells. ( e , f ) Efficient transfection of Orai1-encoding plasmids demonstrated by western blotting for myc-tag. ( g ) LPA-induced Ca 2+ entry was found to be blocked in Orai1 R91W -expressing cells. ( h ) Keratinocytes were cotransfected with Orai1, nuclear factor of activated T cell (NFAT)-directed, and Renilla luciferase plasmids. Cells were then treated with LPA and 1.2 mℳ Ca 2+ and assessed for NFAT transcriptional activity. Overexpression of Orai1 R91W significantly impaired NFAT activation. Data represent mean±SEM; n ⩾3 unless otherwise stated, * P
    Fura Pe3 Am, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Synaptic Systems abeta pe3
    Kinetics of homozygous E8 mice. Immunohistochemical analysis of hom E8 animals from the age of 1–12 months compared to wt animals (A-F) . Staining with 6E10 revealed progressive increase of total <t>Aβ</t> immunoreactivity from the age of 1–9 months with a decline at 12 months (A) . Increase of <t>pE3-Aβ</t> reactivity from 3–9 months and decline at 12 months (B) is accompanied by GFAP reactivity (C) . Apoptotic processes are indicated provided by progressive caspase 3 activity beginning at the age of 3 months (D) . The lateral striatum shows a progressive decrease of DARPP-32 reactivity of neuropil from the age of 6 months (E) . The different processes result in decreased NeuN positive cells of E8 animals indicating cell loss beginning in at the age of 6 months (F) . Quantification of pE3-Aβ positive cells in the striatum of hom E8 animals revealed exponential increase from 3–9 months and declining numbers at the age of 12 months to the level of 6 months (G) ; data were analyzed by one-way ANOVA followed by Newman-Keuls post-hoc test and represent means ± SEM; ** p
    Abeta Pe3, supplied by Synaptic Systems, used in various techniques. Bioz Stars score: 85/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    CoolLED coolled pe3
    Kinetics of homozygous E8 mice. Immunohistochemical analysis of hom E8 animals from the age of 1–12 months compared to wt animals (A-F) . Staining with 6E10 revealed progressive increase of total <t>Aβ</t> immunoreactivity from the age of 1–9 months with a decline at 12 months (A) . Increase of <t>pE3-Aβ</t> reactivity from 3–9 months and decline at 12 months (B) is accompanied by GFAP reactivity (C) . Apoptotic processes are indicated provided by progressive caspase 3 activity beginning at the age of 3 months (D) . The lateral striatum shows a progressive decrease of DARPP-32 reactivity of neuropil from the age of 6 months (E) . The different processes result in decreased NeuN positive cells of E8 animals indicating cell loss beginning in at the age of 6 months (F) . Quantification of pE3-Aβ positive cells in the striatum of hom E8 animals revealed exponential increase from 3–9 months and declining numbers at the age of 12 months to the level of 6 months (G) ; data were analyzed by one-way ANOVA followed by Newman-Keuls post-hoc test and represent means ± SEM; ** p
    Coolled Pe3, supplied by CoolLED, 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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    88
    Thermo Fisher fura pe3
    Effect of ATRA on Ca 2+ influx in A549 cells and TRPC channels. A, A549 cells were loaded with 2 µM <t>Fura-PE3/AM</t> and the Ca 2+ was measured as the ratio (F 340 /F 380 ) of Ca 2+ dye fluorescence. Ca 2+ release was evoked by trypsin (0.2 nM) and Ca 2+ influx was introduced by 1.5 mM Ca 2+ in the perfusion solution. The cells were incubated with 1 µM ATRA for 96 hours and the control cells were incubated with same volume of vehicle. B, Mean ± s.e.m. data for the Ca 2+ entry after the store depletion by trypsin as shown in (A). n = 21–32 for each group, *** P
    Fura Pe3, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Fisher Scientific fura pe3 am ester
    Effect of ATRA on Ca 2+ influx in A549 cells and TRPC channels. A, A549 cells were loaded with 2 µM <t>Fura-PE3/AM</t> and the Ca 2+ was measured as the ratio (F 340 /F 380 ) of Ca 2+ dye fluorescence. Ca 2+ release was evoked by trypsin (0.2 nM) and Ca 2+ influx was introduced by 1.5 mM Ca 2+ in the perfusion solution. The cells were incubated with 1 µM ATRA for 96 hours and the control cells were incubated with same volume of vehicle. B, Mean ± s.e.m. data for the Ca 2+ entry after the store depletion by trypsin as shown in (A). n = 21–32 for each group, *** P
    Fura Pe3 Am Ester, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Synaptic Systems pyro gluabeta pe3
    Effect of ATRA on Ca 2+ influx in A549 cells and TRPC channels. A, A549 cells were loaded with 2 µM <t>Fura-PE3/AM</t> and the Ca 2+ was measured as the ratio (F 340 /F 380 ) of Ca 2+ dye fluorescence. Ca 2+ release was evoked by trypsin (0.2 nM) and Ca 2+ influx was introduced by 1.5 mM Ca 2+ in the perfusion solution. The cells were incubated with 1 µM ATRA for 96 hours and the control cells were incubated with same volume of vehicle. B, Mean ± s.e.m. data for the Ca 2+ entry after the store depletion by trypsin as shown in (A). n = 21–32 for each group, *** P
    Pyro Gluabeta Pe3, supplied by Synaptic Systems, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Thermo Fisher primer express version 3 0 pe3
    Effect of ATRA on Ca 2+ influx in A549 cells and TRPC channels. A, A549 cells were loaded with 2 µM <t>Fura-PE3/AM</t> and the Ca 2+ was measured as the ratio (F 340 /F 380 ) of Ca 2+ dye fluorescence. Ca 2+ release was evoked by trypsin (0.2 nM) and Ca 2+ influx was introduced by 1.5 mM Ca 2+ in the perfusion solution. The cells were incubated with 1 µM ATRA for 96 hours and the control cells were incubated with same volume of vehicle. B, Mean ± s.e.m. data for the Ca 2+ entry after the store depletion by trypsin as shown in (A). n = 21–32 for each group, *** P
    Primer Express Version 3 0 Pe3, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    IDEX Health & Science fura pe3
    Effect of ATRA on Ca 2+ influx in A549 cells and TRPC channels. A, A549 cells were loaded with 2 µM <t>Fura-PE3/AM</t> and the Ca 2+ was measured as the ratio (F 340 /F 380 ) of Ca 2+ dye fluorescence. Ca 2+ release was evoked by trypsin (0.2 nM) and Ca 2+ influx was introduced by 1.5 mM Ca 2+ in the perfusion solution. The cells were incubated with 1 µM ATRA for 96 hours and the control cells were incubated with same volume of vehicle. B, Mean ± s.e.m. data for the Ca 2+ entry after the store depletion by trypsin as shown in (A). n = 21–32 for each group, *** P
    Fura Pe3, supplied by IDEX Health & Science, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    LabCorp fura pe3
    Effect of ATRA on Ca 2+ influx in A549 cells and TRPC channels. A, A549 cells were loaded with 2 µM <t>Fura-PE3/AM</t> and the Ca 2+ was measured as the ratio (F 340 /F 380 ) of Ca 2+ dye fluorescence. Ca 2+ release was evoked by trypsin (0.2 nM) and Ca 2+ influx was introduced by 1.5 mM Ca 2+ in the perfusion solution. The cells were incubated with 1 µM ATRA for 96 hours and the control cells were incubated with same volume of vehicle. B, Mean ± s.e.m. data for the Ca 2+ entry after the store depletion by trypsin as shown in (A). n = 21–32 for each group, *** P
    Fura Pe3, supplied by LabCorp, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore ratiometric dye fura pe3
    Effect of ATRA on Ca 2+ influx in A549 cells and TRPC channels. A, A549 cells were loaded with 2 µM <t>Fura-PE3/AM</t> and the Ca 2+ was measured as the ratio (F 340 /F 380 ) of Ca 2+ dye fluorescence. Ca 2+ release was evoked by trypsin (0.2 nM) and Ca 2+ influx was introduced by 1.5 mM Ca 2+ in the perfusion solution. The cells were incubated with 1 µM ATRA for 96 hours and the control cells were incubated with same volume of vehicle. B, Mean ± s.e.m. data for the Ca 2+ entry after the store depletion by trypsin as shown in (A). n = 21–32 for each group, *** P
    Ratiometric Dye Fura Pe3, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Millipore doxycycline
    Effect of ATRA on Ca 2+ influx in A549 cells and TRPC channels. A, A549 cells were loaded with 2 µM <t>Fura-PE3/AM</t> and the Ca 2+ was measured as the ratio (F 340 /F 380 ) of Ca 2+ dye fluorescence. Ca 2+ release was evoked by trypsin (0.2 nM) and Ca 2+ influx was introduced by 1.5 mM Ca 2+ in the perfusion solution. The cells were incubated with 1 µM ATRA for 96 hours and the control cells were incubated with same volume of vehicle. B, Mean ± s.e.m. data for the Ca 2+ entry after the store depletion by trypsin as shown in (A). n = 21–32 for each group, *** P
    Doxycycline, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 13486 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Dojindo Labs fura pe3 am
    Effect of ATRA on Ca 2+ influx in A549 cells and TRPC channels. A, A549 cells were loaded with 2 µM <t>Fura-PE3/AM</t> and the Ca 2+ was measured as the ratio (F 340 /F 380 ) of Ca 2+ dye fluorescence. Ca 2+ release was evoked by trypsin (0.2 nM) and Ca 2+ influx was introduced by 1.5 mM Ca 2+ in the perfusion solution. The cells were incubated with 1 µM ATRA for 96 hours and the control cells were incubated with same volume of vehicle. B, Mean ± s.e.m. data for the Ca 2+ entry after the store depletion by trypsin as shown in (A). n = 21–32 for each group, *** P
    Fura Pe3 Am, supplied by Dojindo Labs, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    FUJIFILM fura pe3 am
    Effect of ATRA on Ca 2+ influx in A549 cells and TRPC channels. A, A549 cells were loaded with 2 µM <t>Fura-PE3/AM</t> and the Ca 2+ was measured as the ratio (F 340 /F 380 ) of Ca 2+ dye fluorescence. Ca 2+ release was evoked by trypsin (0.2 nM) and Ca 2+ influx was introduced by 1.5 mM Ca 2+ in the perfusion solution. The cells were incubated with 1 µM ATRA for 96 hours and the control cells were incubated with same volume of vehicle. B, Mean ± s.e.m. data for the Ca 2+ entry after the store depletion by trypsin as shown in (A). n = 21–32 for each group, *** P
    Fura Pe3 Am, supplied by FUJIFILM, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Scientific Commodities polyethylene pe3 tubing
    Effect of ATRA on Ca 2+ influx in A549 cells and TRPC channels. A, A549 cells were loaded with 2 µM <t>Fura-PE3/AM</t> and the Ca 2+ was measured as the ratio (F 340 /F 380 ) of Ca 2+ dye fluorescence. Ca 2+ release was evoked by trypsin (0.2 nM) and Ca 2+ influx was introduced by 1.5 mM Ca 2+ in the perfusion solution. The cells were incubated with 1 µM ATRA for 96 hours and the control cells were incubated with same volume of vehicle. B, Mean ± s.e.m. data for the Ca 2+ entry after the store depletion by trypsin as shown in (A). n = 21–32 for each group, *** P
    Polyethylene Pe3 Tubing, supplied by Scientific Commodities, used in various techniques. Bioz Stars score: 86/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Scientific Commodities medical grade polyethylene pe3 tubings
    Effect of ATRA on Ca 2+ influx in A549 cells and TRPC channels. A, A549 cells were loaded with 2 µM <t>Fura-PE3/AM</t> and the Ca 2+ was measured as the ratio (F 340 /F 380 ) of Ca 2+ dye fluorescence. Ca 2+ release was evoked by trypsin (0.2 nM) and Ca 2+ influx was introduced by 1.5 mM Ca 2+ in the perfusion solution. The cells were incubated with 1 µM ATRA for 96 hours and the control cells were incubated with same volume of vehicle. B, Mean ± s.e.m. data for the Ca 2+ entry after the store depletion by trypsin as shown in (A). n = 21–32 for each group, *** P
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    Millipore ca2 indicator fura pe3
    Effect of ATRA on Ca 2+ influx in A549 cells and TRPC channels. A, A549 cells were loaded with 2 µM <t>Fura-PE3/AM</t> and the Ca 2+ was measured as the ratio (F 340 /F 380 ) of Ca 2+ dye fluorescence. Ca 2+ release was evoked by trypsin (0.2 nM) and Ca 2+ influx was introduced by 1.5 mM Ca 2+ in the perfusion solution. The cells were incubated with 1 µM ATRA for 96 hours and the control cells were incubated with same volume of vehicle. B, Mean ± s.e.m. data for the Ca 2+ entry after the store depletion by trypsin as shown in (A). n = 21–32 for each group, *** P
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    Image Search Results


    Effect of La 3+ on HPV in IPA A and B , tension and fura PE-3 F 340/380 fluorescence ratio, respectively, measured simultaneously in a single IPA under control conditions and in the presence of 1 and 100 μM La 3+ . C and D , mean results for five IPAs: control, ○; 1 μM La 3+ , •; and 100 μM La 3+ , ▪. * P

    Journal: The Journal of Physiology

    Article Title: Voltage-independent calcium entry in hypoxic pulmonary vasoconstriction of intrapulmonary arteries of the rat

    doi: 10.1111/j.1469-7793.2000.t01-1-00669.x

    Figure Lengend Snippet: Effect of La 3+ on HPV in IPA A and B , tension and fura PE-3 F 340/380 fluorescence ratio, respectively, measured simultaneously in a single IPA under control conditions and in the presence of 1 and 100 μM La 3+ . C and D , mean results for five IPAs: control, ○; 1 μM La 3+ , •; and 100 μM La 3+ , ▪. * P

    Article Snippet: IPAs were loaded with the Ca2+ -sensitive fluorophore fura PE-3, via incubation of the vessels with the acetoxymethyl ester of fura PE-3 (3 μM) for 2 h at room temperature (Sigma-Aldrich Ltd).

    Techniques: Indirect Immunoperoxidase Assay, Fluorescence

    Verapamil and diltiazem have no effect upon either the sustained increase in tension or [Ca 2+ ] i during HPV in polarized or depolarized arteries A and B, IPAs were exposed to hypoxia in the presence of 3 μM PGF 2α (○) and following preincubation with 10 μM verapamil (•); the PGF 2α concentration was raised to 3.5 or 4 μM in the presence of verapamil to match pretone to the control level. A shows tension development in eight IPAs; B depicts fura PE-3 F 340/380 fluorescence ratio, an indication of [Ca 2+ ] i , measured simultaneously in four of these arteries ( * P

    Journal: The Journal of Physiology

    Article Title: Voltage-independent calcium entry in hypoxic pulmonary vasoconstriction of intrapulmonary arteries of the rat

    doi: 10.1111/j.1469-7793.2000.t01-1-00669.x

    Figure Lengend Snippet: Verapamil and diltiazem have no effect upon either the sustained increase in tension or [Ca 2+ ] i during HPV in polarized or depolarized arteries A and B, IPAs were exposed to hypoxia in the presence of 3 μM PGF 2α (○) and following preincubation with 10 μM verapamil (•); the PGF 2α concentration was raised to 3.5 or 4 μM in the presence of verapamil to match pretone to the control level. A shows tension development in eight IPAs; B depicts fura PE-3 F 340/380 fluorescence ratio, an indication of [Ca 2+ ] i , measured simultaneously in four of these arteries ( * P

    Article Snippet: IPAs were loaded with the Ca2+ -sensitive fluorophore fura PE-3, via incubation of the vessels with the acetoxymethyl ester of fura PE-3 (3 μM) for 2 h at room temperature (Sigma-Aldrich Ltd).

    Techniques: Concentration Assay, Fluorescence

    In a therapeutic study initiated well after the onset of cerebral Aβ deposition and gliosis, weekly passive immunization with anti-pE3-Aβ mAb07/1 in 23-monthold APPswe/PS1ΔE9 mice for 7 weeks resulted in the attenuation of pE3-Aβ and general Aβ (R1282 IR) deposition as well as fibrillar amyloid (Thioflavin S) in the hippocampus ( a , b ) and cerebellum ( c , d ) compared to PBS control mice. Immunohistochemical results ( a , c ) and Thioflavin S labeling were quantified by image analysis ( b , d ). Absolute values are provided in table 1. Scale bars, 200 µm. p values: * p > 0.05; *** p > 0.001; n.s. = nonsignificant (p = 0.11); p = 0.06 (strong trend).

    Journal: Neuro-Degenerative Diseases

    Article Title: Passive Immunization against Pyroglutamate-3 Amyloid-? Reduces Plaque Burden in Alzheimer-Like Transgenic Mice: A Pilot Study

    doi: 10.1159/000335913

    Figure Lengend Snippet: In a therapeutic study initiated well after the onset of cerebral Aβ deposition and gliosis, weekly passive immunization with anti-pE3-Aβ mAb07/1 in 23-monthold APPswe/PS1ΔE9 mice for 7 weeks resulted in the attenuation of pE3-Aβ and general Aβ (R1282 IR) deposition as well as fibrillar amyloid (Thioflavin S) in the hippocampus ( a , b ) and cerebellum ( c , d ) compared to PBS control mice. Immunohistochemical results ( a , c ) and Thioflavin S labeling were quantified by image analysis ( b , d ). Absolute values are provided in table 1. Scale bars, 200 µm. p values: * p > 0.05; *** p > 0.001; n.s. = nonsignificant (p = 0.11); p = 0.06 (strong trend).

    Article Snippet: Mice were vaccinated weekly by intraperitoneal injection of 200 μg of a new mouse IgG1 mAb specific for pE3-Aβ [mAb07/1; Probiodrug AG, Halle (Saale), Germany] or 100 μl of PBS (as a control).

    Techniques: Mouse Assay, Immunohistochemistry, Labeling

    In a prevention study initiated during the early stage of plaque deposition, weekly passive immunization of APPswe/PS1ΔlE9 mice with anti-pE3-Aβ mAb07/1 from 5.8 to 13.8 months of age significantly reduced pE3-Aβ as well as general Aβ (R1282 IR) and fibrillar amyloid (Thioflavin S) deposition in the hippocampus ( a , b ) and cerebellum ( c , d ) compared to that in PBS control mice. Immunohistochemical results ( a , c ) and Thioflavin S labeling were quantified by image analysis ( b , d ). Absolute values are provided in table 1. Scale bars, 200 µm. p values: * p > 0.05; ** p > 0.01; *** p > 0.001; n.s. = nonsignificant (p = 0.089).

    Journal: Neuro-Degenerative Diseases

    Article Title: Passive Immunization against Pyroglutamate-3 Amyloid-? Reduces Plaque Burden in Alzheimer-Like Transgenic Mice: A Pilot Study

    doi: 10.1159/000335913

    Figure Lengend Snippet: In a prevention study initiated during the early stage of plaque deposition, weekly passive immunization of APPswe/PS1ΔlE9 mice with anti-pE3-Aβ mAb07/1 from 5.8 to 13.8 months of age significantly reduced pE3-Aβ as well as general Aβ (R1282 IR) and fibrillar amyloid (Thioflavin S) deposition in the hippocampus ( a , b ) and cerebellum ( c , d ) compared to that in PBS control mice. Immunohistochemical results ( a , c ) and Thioflavin S labeling were quantified by image analysis ( b , d ). Absolute values are provided in table 1. Scale bars, 200 µm. p values: * p > 0.05; ** p > 0.01; *** p > 0.001; n.s. = nonsignificant (p = 0.089).

    Article Snippet: Mice were vaccinated weekly by intraperitoneal injection of 200 μg of a new mouse IgG1 mAb specific for pE3-Aβ [mAb07/1; Probiodrug AG, Halle (Saale), Germany] or 100 μl of PBS (as a control).

    Techniques: Mouse Assay, Immunohistochemistry, Labeling

    Characterization of pE3-Aβ mAb07/1. a Western blot of Aβ(1–40), Aβ(3–40) and pE3-Aβ(3–40), detected with mAb07/1 or mAb6E10. Peptides (20 ng each) were separated in gels containing 8 M urea. The anti-pE3-Aβ mAb does not detect the truncated precursor or full-length Aβ. b Analysis of antibody binding using surface plasmon resonance. The peptides were covalently linked and the antibody applied in a buffered solution. Significant binding of mAb07/1 was only observed with pE3-Aβ (3–40) immobilized on the surface. Other peptides analyzed are: MCP-1 and 2; gastrin; GnRH; neurotensin; orexin; TRH; the N-terminus of collagen, and fibronectin.

    Journal: Neuro-Degenerative Diseases

    Article Title: Passive Immunization against Pyroglutamate-3 Amyloid-? Reduces Plaque Burden in Alzheimer-Like Transgenic Mice: A Pilot Study

    doi: 10.1159/000335913

    Figure Lengend Snippet: Characterization of pE3-Aβ mAb07/1. a Western blot of Aβ(1–40), Aβ(3–40) and pE3-Aβ(3–40), detected with mAb07/1 or mAb6E10. Peptides (20 ng each) were separated in gels containing 8 M urea. The anti-pE3-Aβ mAb does not detect the truncated precursor or full-length Aβ. b Analysis of antibody binding using surface plasmon resonance. The peptides were covalently linked and the antibody applied in a buffered solution. Significant binding of mAb07/1 was only observed with pE3-Aβ (3–40) immobilized on the surface. Other peptides analyzed are: MCP-1 and 2; gastrin; GnRH; neurotensin; orexin; TRH; the N-terminus of collagen, and fibronectin.

    Article Snippet: Mice were vaccinated weekly by intraperitoneal injection of 200 μg of a new mouse IgG1 mAb specific for pE3-Aβ [mAb07/1; Probiodrug AG, Halle (Saale), Germany] or 100 μl of PBS (as a control).

    Techniques: Western Blot, Binding Assay, SPR Assay

    Effect of prolonged exposure to insulin on [Ca 2+ ] i , total calcium current (I Ca 2+ ), and SERCA2 protein. (A and B) INS1E β -cells cultured on glass coverslips were loaded with fura-2–PE3 (2 µmol/L) and Mag–fluo-4-AM (1 μM), respectively (Molecular Probes, Eugene, OR). Pluronic acid (0.01%) was added to the medium and cells were incubated for 30 min at 37°C. [Ca 2+ ] i was measured using an Olympus IX-50 inverted epifluorescence microscope (Olympus, Tokyo, Japan). Absolute [Ca 2+ ] i was determined from R of Ca 2+ -bound fura-2 (excited at 340 nm) to unbound fura-2 (excited at 380 nM). The Ca 2+ levels were determined using a standard equation for calibration. R max and R min were obtained by adding ionomycin (10 µM) or EGTA (1 mM), respectively, to fura-2–loaded β -cells at the end of each experiment. [Ca 2+ ] i was obtained using an excitation wavelength of 488 nm and an emission band-pass filter of 515/15 nm. Means ± SEM are from four independent experiments. The I Ca 2+ (Ba 2+ ) from control and insulin pretreated cells for 48 h were evoked by a 100 msec step depolarization under whole-cell voltage clamp configuration. * P

    Journal: Journal of the Endocrine Society

    Article Title: Prolonged Exposure to Insulin Inactivates Akt and Erk1/2 and Increases Pancreatic Islet and INS1E β-Cell Apoptosis

    doi: 10.1210/js.2018-00140

    Figure Lengend Snippet: Effect of prolonged exposure to insulin on [Ca 2+ ] i , total calcium current (I Ca 2+ ), and SERCA2 protein. (A and B) INS1E β -cells cultured on glass coverslips were loaded with fura-2–PE3 (2 µmol/L) and Mag–fluo-4-AM (1 μM), respectively (Molecular Probes, Eugene, OR). Pluronic acid (0.01%) was added to the medium and cells were incubated for 30 min at 37°C. [Ca 2+ ] i was measured using an Olympus IX-50 inverted epifluorescence microscope (Olympus, Tokyo, Japan). Absolute [Ca 2+ ] i was determined from R of Ca 2+ -bound fura-2 (excited at 340 nm) to unbound fura-2 (excited at 380 nM). The Ca 2+ levels were determined using a standard equation for calibration. R max and R min were obtained by adding ionomycin (10 µM) or EGTA (1 mM), respectively, to fura-2–loaded β -cells at the end of each experiment. [Ca 2+ ] i was obtained using an excitation wavelength of 488 nm and an emission band-pass filter of 515/15 nm. Means ± SEM are from four independent experiments. The I Ca 2+ (Ba 2+ ) from control and insulin pretreated cells for 48 h were evoked by a 100 msec step depolarization under whole-cell voltage clamp configuration. * P

    Article Snippet: For intracellular Ca2+ concentration ([Ca2+ ]i ) measurement, cells were loaded with fura-2–PE3 (2 µmol/L) (Molecular Probes, Eugene, OR).

    Techniques: Cell Culture, Incubation, Inverted Epifluorescence

    Lysophosphatidic acid (LPA)-induced Ca 2+ entry requires sufficient levels of STIM1 and functional Orai1. ( a – c ) Relative efficiency of small interfering RNA (siRNA)-mediated STIM1 silencing compared to scrambled (scr) control by quantitative PCR and western blotting. ( e ) Fura-PE3-based Ca 2+ imaging showed impaired LPA-induced Ca 2+ entry in STIM1-knockdown cells. ( e , f ) Efficient transfection of Orai1-encoding plasmids demonstrated by western blotting for myc-tag. ( g ) LPA-induced Ca 2+ entry was found to be blocked in Orai1 R91W -expressing cells. ( h ) Keratinocytes were cotransfected with Orai1, nuclear factor of activated T cell (NFAT)-directed, and Renilla luciferase plasmids. Cells were then treated with LPA and 1.2 mℳ Ca 2+ and assessed for NFAT transcriptional activity. Overexpression of Orai1 R91W significantly impaired NFAT activation. Data represent mean±SEM; n ⩾3 unless otherwise stated, * P

    Journal: The Journal of Investigative Dermatology

    Article Title: Lysophosphatidic Acid Promotes Cell Migration through STIM1- and Orai1-Mediated Ca2+i Mobilization and NFAT2 Activation

    doi: 10.1038/jid.2012.370

    Figure Lengend Snippet: Lysophosphatidic acid (LPA)-induced Ca 2+ entry requires sufficient levels of STIM1 and functional Orai1. ( a – c ) Relative efficiency of small interfering RNA (siRNA)-mediated STIM1 silencing compared to scrambled (scr) control by quantitative PCR and western blotting. ( e ) Fura-PE3-based Ca 2+ imaging showed impaired LPA-induced Ca 2+ entry in STIM1-knockdown cells. ( e , f ) Efficient transfection of Orai1-encoding plasmids demonstrated by western blotting for myc-tag. ( g ) LPA-induced Ca 2+ entry was found to be blocked in Orai1 R91W -expressing cells. ( h ) Keratinocytes were cotransfected with Orai1, nuclear factor of activated T cell (NFAT)-directed, and Renilla luciferase plasmids. Cells were then treated with LPA and 1.2 mℳ Ca 2+ and assessed for NFAT transcriptional activity. Overexpression of Orai1 R91W significantly impaired NFAT activation. Data represent mean±SEM; n ⩾3 unless otherwise stated, * P

    Article Snippet: Ca2+ imaging Keratinocytes seeded in Willco glass-bottomed dishes (Intracel, Royston, UK) were subjected to Ca2+ i imaging using Fluo-4-AM (Invitrogen) or Fura-PE3-AM (Calbiochem) as described ( ).

    Techniques: Functional Assay, Small Interfering RNA, Real-time Polymerase Chain Reaction, Western Blot, Imaging, Transfection, Expressing, Luciferase, Activity Assay, Over Expression, Activation Assay

    Kinetics of homozygous E8 mice. Immunohistochemical analysis of hom E8 animals from the age of 1–12 months compared to wt animals (A-F) . Staining with 6E10 revealed progressive increase of total Aβ immunoreactivity from the age of 1–9 months with a decline at 12 months (A) . Increase of pE3-Aβ reactivity from 3–9 months and decline at 12 months (B) is accompanied by GFAP reactivity (C) . Apoptotic processes are indicated provided by progressive caspase 3 activity beginning at the age of 3 months (D) . The lateral striatum shows a progressive decrease of DARPP-32 reactivity of neuropil from the age of 6 months (E) . The different processes result in decreased NeuN positive cells of E8 animals indicating cell loss beginning in at the age of 6 months (F) . Quantification of pE3-Aβ positive cells in the striatum of hom E8 animals revealed exponential increase from 3–9 months and declining numbers at the age of 12 months to the level of 6 months (G) ; data were analyzed by one-way ANOVA followed by Newman-Keuls post-hoc test and represent means ± SEM; ** p

    Journal: BMC Neuroscience

    Article Title: Glutaminyl cyclase-mediated toxicity of pyroglutamate-beta amyloid induces striatal neurodegeneration

    doi: 10.1186/1471-2202-14-108

    Figure Lengend Snippet: Kinetics of homozygous E8 mice. Immunohistochemical analysis of hom E8 animals from the age of 1–12 months compared to wt animals (A-F) . Staining with 6E10 revealed progressive increase of total Aβ immunoreactivity from the age of 1–9 months with a decline at 12 months (A) . Increase of pE3-Aβ reactivity from 3–9 months and decline at 12 months (B) is accompanied by GFAP reactivity (C) . Apoptotic processes are indicated provided by progressive caspase 3 activity beginning at the age of 3 months (D) . The lateral striatum shows a progressive decrease of DARPP-32 reactivity of neuropil from the age of 6 months (E) . The different processes result in decreased NeuN positive cells of E8 animals indicating cell loss beginning in at the age of 6 months (F) . Quantification of pE3-Aβ positive cells in the striatum of hom E8 animals revealed exponential increase from 3–9 months and declining numbers at the age of 12 months to the level of 6 months (G) ; data were analyzed by one-way ANOVA followed by Newman-Keuls post-hoc test and represent means ± SEM; ** p

    Article Snippet: In this study Aβ-specific antibody 6E10 (mouse monoclonal; SIG-39320; Covance; 1:10,000 dilution), pE3-Aβ-specific antibodies Abeta-pE3 (rabbit polyclonal; 218003; Synaptic Systems; 1:100,000 dilution) and Abeta-pE3 (mouse monoclonal; 218011; Synaptic Systems; 1:10,000 dilution), QC-specific antibody 1302 (rabbit polyclonal; Probiodrug; 1:1,000 dilution), and human QC-specific antibody 8696 (rabbit polyclonal, Probiodrug; 1:1,000 dilution), glia-specific antibody GFAP (rabbit polyclonal, Z0334; DAKOCytomation; 1:30,000 dilution), activated caspase 3-specific antibody (rabbit polyclonal, 9661, Cell Signaling Technology; dilution 1:100), neuron-specific antibody NeuN (mouse monoclonal; MAB377; Chemicon-Millipore; 1:10,000 dilution), and DARPP32-specific antibody (rabbit monoclonal; 1710–1; Epitomics; 1:10,000 dilution) were used as primary antibodies.

    Techniques: Mouse Assay, Immunohistochemistry, Staining, Activity Assay

    Influence of QC overexpression on pE3-Aβ formation. Staining of striatal sections of transgenic E8-hQC mice demonstrated an increase of pE3-Aβ positive cells by het and hom hQC overexpression, respectively at the age of 3 months (A) and 6 months (B) . Quantification of these pE3-Aβ positive cells revealed a significant increase for het and hom overexpression of hQC in homs E8 animals at the age of 6 months and in het/het E85 animals at the age of 7 months (C) ; E8 were analyzed by one-way ANOVA followed by Newman-Keuls post-hoc test; E85 were analyzed by unpaired t-test; data represent means ± SEM; * p

    Journal: BMC Neuroscience

    Article Title: Glutaminyl cyclase-mediated toxicity of pyroglutamate-beta amyloid induces striatal neurodegeneration

    doi: 10.1186/1471-2202-14-108

    Figure Lengend Snippet: Influence of QC overexpression on pE3-Aβ formation. Staining of striatal sections of transgenic E8-hQC mice demonstrated an increase of pE3-Aβ positive cells by het and hom hQC overexpression, respectively at the age of 3 months (A) and 6 months (B) . Quantification of these pE3-Aβ positive cells revealed a significant increase for het and hom overexpression of hQC in homs E8 animals at the age of 6 months and in het/het E85 animals at the age of 7 months (C) ; E8 were analyzed by one-way ANOVA followed by Newman-Keuls post-hoc test; E85 were analyzed by unpaired t-test; data represent means ± SEM; * p

    Article Snippet: In this study Aβ-specific antibody 6E10 (mouse monoclonal; SIG-39320; Covance; 1:10,000 dilution), pE3-Aβ-specific antibodies Abeta-pE3 (rabbit polyclonal; 218003; Synaptic Systems; 1:100,000 dilution) and Abeta-pE3 (mouse monoclonal; 218011; Synaptic Systems; 1:10,000 dilution), QC-specific antibody 1302 (rabbit polyclonal; Probiodrug; 1:1,000 dilution), and human QC-specific antibody 8696 (rabbit polyclonal, Probiodrug; 1:1,000 dilution), glia-specific antibody GFAP (rabbit polyclonal, Z0334; DAKOCytomation; 1:30,000 dilution), activated caspase 3-specific antibody (rabbit polyclonal, 9661, Cell Signaling Technology; dilution 1:100), neuron-specific antibody NeuN (mouse monoclonal; MAB377; Chemicon-Millipore; 1:10,000 dilution), and DARPP32-specific antibody (rabbit monoclonal; 1710–1; Epitomics; 1:10,000 dilution) were used as primary antibodies.

    Techniques: Over Expression, Staining, Transgenic Assay, Mouse Assay

    Striatal (co-)localizations of Aβ, pE3-Aβ, QC and astroglia in lateral striatum. Double immunofluorescent labeling of total Aβ (red) and pE3-Aβ (green) revealed intraneuronal colocalization of tg product and pE-modified Aβ in the lateral striatum of hom E8 animals (A) . Double immunofluorescent labeling of the lateral striatum showed mQC-specific immunoreactivity (red), which is colocalized with pE3-Aβ (green) in hom E8 animals (B) . Double immunofluorescent labeling revealed increased GFAP reactivity (green) of activated astroglia in the vicinity of pE3-Aβ positive cells (red) in the lateral striatum of hom E8 animals (C) .

    Journal: BMC Neuroscience

    Article Title: Glutaminyl cyclase-mediated toxicity of pyroglutamate-beta amyloid induces striatal neurodegeneration

    doi: 10.1186/1471-2202-14-108

    Figure Lengend Snippet: Striatal (co-)localizations of Aβ, pE3-Aβ, QC and astroglia in lateral striatum. Double immunofluorescent labeling of total Aβ (red) and pE3-Aβ (green) revealed intraneuronal colocalization of tg product and pE-modified Aβ in the lateral striatum of hom E8 animals (A) . Double immunofluorescent labeling of the lateral striatum showed mQC-specific immunoreactivity (red), which is colocalized with pE3-Aβ (green) in hom E8 animals (B) . Double immunofluorescent labeling revealed increased GFAP reactivity (green) of activated astroglia in the vicinity of pE3-Aβ positive cells (red) in the lateral striatum of hom E8 animals (C) .

    Article Snippet: In this study Aβ-specific antibody 6E10 (mouse monoclonal; SIG-39320; Covance; 1:10,000 dilution), pE3-Aβ-specific antibodies Abeta-pE3 (rabbit polyclonal; 218003; Synaptic Systems; 1:100,000 dilution) and Abeta-pE3 (mouse monoclonal; 218011; Synaptic Systems; 1:10,000 dilution), QC-specific antibody 1302 (rabbit polyclonal; Probiodrug; 1:1,000 dilution), and human QC-specific antibody 8696 (rabbit polyclonal, Probiodrug; 1:1,000 dilution), glia-specific antibody GFAP (rabbit polyclonal, Z0334; DAKOCytomation; 1:30,000 dilution), activated caspase 3-specific antibody (rabbit polyclonal, 9661, Cell Signaling Technology; dilution 1:100), neuron-specific antibody NeuN (mouse monoclonal; MAB377; Chemicon-Millipore; 1:10,000 dilution), and DARPP32-specific antibody (rabbit monoclonal; 1710–1; Epitomics; 1:10,000 dilution) were used as primary antibodies.

    Techniques: Labeling, Modification

    QC overexpression. Double immunofluorescent labeling of hom/hom E8-hQC animals revealed human QC specific immunoreactivity (red) colocalized with pE3-Aβ (green) in the striatum (A) . Analysis of QC activity at the age of 6 months revealed an indistinguishable activity for ETNA animals compared to wt animals and a ~60-fold increase for het and ~100-fold increase for hom overexpression of hQC (B) ; data were analyzed by one-way ANOVA followed by Newman-Keuls post-hoc test and represent means ± SEM; *** p

    Journal: BMC Neuroscience

    Article Title: Glutaminyl cyclase-mediated toxicity of pyroglutamate-beta amyloid induces striatal neurodegeneration

    doi: 10.1186/1471-2202-14-108

    Figure Lengend Snippet: QC overexpression. Double immunofluorescent labeling of hom/hom E8-hQC animals revealed human QC specific immunoreactivity (red) colocalized with pE3-Aβ (green) in the striatum (A) . Analysis of QC activity at the age of 6 months revealed an indistinguishable activity for ETNA animals compared to wt animals and a ~60-fold increase for het and ~100-fold increase for hom overexpression of hQC (B) ; data were analyzed by one-way ANOVA followed by Newman-Keuls post-hoc test and represent means ± SEM; *** p

    Article Snippet: In this study Aβ-specific antibody 6E10 (mouse monoclonal; SIG-39320; Covance; 1:10,000 dilution), pE3-Aβ-specific antibodies Abeta-pE3 (rabbit polyclonal; 218003; Synaptic Systems; 1:100,000 dilution) and Abeta-pE3 (mouse monoclonal; 218011; Synaptic Systems; 1:10,000 dilution), QC-specific antibody 1302 (rabbit polyclonal; Probiodrug; 1:1,000 dilution), and human QC-specific antibody 8696 (rabbit polyclonal, Probiodrug; 1:1,000 dilution), glia-specific antibody GFAP (rabbit polyclonal, Z0334; DAKOCytomation; 1:30,000 dilution), activated caspase 3-specific antibody (rabbit polyclonal, 9661, Cell Signaling Technology; dilution 1:100), neuron-specific antibody NeuN (mouse monoclonal; MAB377; Chemicon-Millipore; 1:10,000 dilution), and DARPP32-specific antibody (rabbit monoclonal; 1710–1; Epitomics; 1:10,000 dilution) were used as primary antibodies.

    Techniques: Over Expression, Labeling, Activity Assay

    Construct expression and processing. In the construct of ETNA Aβ(3–42) is fused to pre-pro-TRH for product liberation within the secretory pathway. After prohormone convertase cleavage the N-terminally truncated Aβ peptide is transported into the trans-Golgi and secretory vesicles, where the N-terminus is available to QC for cyclization (A) ; adapted from Alexandru et al., 2011. Quantification of protein levels by ELISA of ETNA brains revealed highest total Aβ levels for hom E8, lowest for hom E5 and intermediate for het/het E85 at two different ages (B) ; total Aβ values expressed as percentage of hom E8 at the age of 9 months; data represent means ± SEM; n ≥ 5 animals per genotype. These protein level relations between the three lines are reflected by similar relations of pE3-Aβ levels at the age of 9 months (C) ; data represent means ± SEM; n ≥ 5 animals per genotype.

    Journal: BMC Neuroscience

    Article Title: Glutaminyl cyclase-mediated toxicity of pyroglutamate-beta amyloid induces striatal neurodegeneration

    doi: 10.1186/1471-2202-14-108

    Figure Lengend Snippet: Construct expression and processing. In the construct of ETNA Aβ(3–42) is fused to pre-pro-TRH for product liberation within the secretory pathway. After prohormone convertase cleavage the N-terminally truncated Aβ peptide is transported into the trans-Golgi and secretory vesicles, where the N-terminus is available to QC for cyclization (A) ; adapted from Alexandru et al., 2011. Quantification of protein levels by ELISA of ETNA brains revealed highest total Aβ levels for hom E8, lowest for hom E5 and intermediate for het/het E85 at two different ages (B) ; total Aβ values expressed as percentage of hom E8 at the age of 9 months; data represent means ± SEM; n ≥ 5 animals per genotype. These protein level relations between the three lines are reflected by similar relations of pE3-Aβ levels at the age of 9 months (C) ; data represent means ± SEM; n ≥ 5 animals per genotype.

    Article Snippet: In this study Aβ-specific antibody 6E10 (mouse monoclonal; SIG-39320; Covance; 1:10,000 dilution), pE3-Aβ-specific antibodies Abeta-pE3 (rabbit polyclonal; 218003; Synaptic Systems; 1:100,000 dilution) and Abeta-pE3 (mouse monoclonal; 218011; Synaptic Systems; 1:10,000 dilution), QC-specific antibody 1302 (rabbit polyclonal; Probiodrug; 1:1,000 dilution), and human QC-specific antibody 8696 (rabbit polyclonal, Probiodrug; 1:1,000 dilution), glia-specific antibody GFAP (rabbit polyclonal, Z0334; DAKOCytomation; 1:30,000 dilution), activated caspase 3-specific antibody (rabbit polyclonal, 9661, Cell Signaling Technology; dilution 1:100), neuron-specific antibody NeuN (mouse monoclonal; MAB377; Chemicon-Millipore; 1:10,000 dilution), and DARPP32-specific antibody (rabbit monoclonal; 1710–1; Epitomics; 1:10,000 dilution) were used as primary antibodies.

    Techniques: Construct, Expressing, Enzyme-linked Immunosorbent Assay

    Effect of ATRA on Ca 2+ influx in A549 cells and TRPC channels. A, A549 cells were loaded with 2 µM Fura-PE3/AM and the Ca 2+ was measured as the ratio (F 340 /F 380 ) of Ca 2+ dye fluorescence. Ca 2+ release was evoked by trypsin (0.2 nM) and Ca 2+ influx was introduced by 1.5 mM Ca 2+ in the perfusion solution. The cells were incubated with 1 µM ATRA for 96 hours and the control cells were incubated with same volume of vehicle. B, Mean ± s.e.m. data for the Ca 2+ entry after the store depletion by trypsin as shown in (A). n = 21–32 for each group, *** P

    Journal: PLoS ONE

    Article Title: Involvement of TRPC Channels in Lung Cancer Cell Differentiation and the Correlation Analysis in Human Non-Small Cell Lung Cancer

    doi: 10.1371/journal.pone.0067637

    Figure Lengend Snippet: Effect of ATRA on Ca 2+ influx in A549 cells and TRPC channels. A, A549 cells were loaded with 2 µM Fura-PE3/AM and the Ca 2+ was measured as the ratio (F 340 /F 380 ) of Ca 2+ dye fluorescence. Ca 2+ release was evoked by trypsin (0.2 nM) and Ca 2+ influx was introduced by 1.5 mM Ca 2+ in the perfusion solution. The cells were incubated with 1 µM ATRA for 96 hours and the control cells were incubated with same volume of vehicle. B, Mean ± s.e.m. data for the Ca 2+ entry after the store depletion by trypsin as shown in (A). n = 21–32 for each group, *** P

    Article Snippet: Gadolinium chloride (Gd3+ ), 2-aminoethoxydiphenyl borate (2-APB), trypsin, all-trans retinoic acid (ATRA), and PCR primers were purchased from Sigma-Aldrich, and Fura-PE3 AM was from Invitrogen (Paisley, UK).

    Techniques: Fluorescence, Incubation