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  • 99
    Synaptic Systems cav2 3
    Representative images and quantification of western blot analysis of calcium- and voltage-gated potassium channels. (A) Total expression of BK and SK2 potassium channels in mouse hippocampus showed no significant differences between WT and <t>Cav2.3-KO</t> mice (WT = 8, KO = 6; p > 0.05). In contrast, in the Cav2.3-KO, the total level of Kv4.2 was significantly increased compared to the WT (WT = 8, KO = 6; p
    Cav2 3, supplied by Synaptic Systems, used in various techniques. Bioz Stars score: 99/100, based on 47 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher hanks balanced salt solution hbss
    Representative images and quantification of western blot analysis of calcium- and voltage-gated potassium channels. (A) Total expression of BK and SK2 potassium channels in mouse hippocampus showed no significant differences between WT and <t>Cav2.3-KO</t> mice (WT = 8, KO = 6; p > 0.05). In contrast, in the Cav2.3-KO, the total level of Kv4.2 was significantly increased compared to the WT (WT = 8, KO = 6; p
    Hanks Balanced Salt Solution Hbss, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2917 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher ca2
    Representative images and quantification of western blot analysis of calcium- and voltage-gated potassium channels. (A) Total expression of BK and SK2 potassium channels in mouse hippocampus showed no significant differences between WT and <t>Cav2.3-KO</t> mice (WT = 8, KO = 6; p > 0.05). In contrast, in the Cav2.3-KO, the total level of Kv4.2 was significantly increased compared to the WT (WT = 8, KO = 6; p
    Ca2, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 5278 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Ipsen Group ca2
    Stimulus-induced secretion consists of CIVDS and CDS components in the somata of DRG neurons. ( A ) Depolarization- and UV-flash-induced C m signals in a DRG neuron dialyzed with 5 mM NP-EGTA and 0.2 mM fura-6F. In <t>Ca2+-free</t> extracellular solution ( left panel ), a step depolarization (200 ms) evoked CIVDS ( C m0 ). When extracellular Ca 2+ was added ( right panel ), UV flashes ( arrows ) evoked a substantial increase of C m ( C m (UV)), which was caused by the Ca 2+ increase in the absence of membrane depolarization. Series conductance ( G s) and membrane current ( I m ), and Ca 2+ trace ([Ca 2+ ]i) are also shown. ( B ) Depolarization-induced C m signals. In Ca 2+ -free extracellular solution ( left panel ), a step depolarization (200 ms) evoked a C m increase (CIVDS) followed by a rapid reversal as in A . C m1 refers to the entire course. Subsequently, extracellular Ca 2+ (2.5 mM) was puffed ( right panel ), and the same stimulus evoked a biphasic response ( C m2 ). The first Cm increase was followed by a small but rapid reversal and then a slow but substantial increase. C m2 − C m1 ( dashed line ) indicated the CDS component.
    Ca2, supplied by Ipsen Group, 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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    99
    Thermo Fisher ca2 indicator fluo 4
    Mast cell degranulation by the GBS pigment requires <t>Ca2</t> + influx ( A ) PCMCs were pretreated with the calcium indicator Fluo-4-AM, and calcium influx was recorded by flow cytometry. At 60 s, either 5 μM A23187 (top panel), 0.5 μM GBS pigment (bottom panel), or an equivalent amount of control Δ cylE extract (middle panel) was added. Mean fluorescence intensities of mast cells before treatment (red) and after treatment (green) are shown. Time is given in seconds. A representative image from one of three independent experiments is shown. ( B ) PCMCs were pretreated with either EGTA (4 mM) or LY294002 (100 μM) for 30 min or with pertussis toxin (PT; 200 ng/ml) for 2 hours. Untreated PCMCs were included as controls for both pretreatment conditions. Subsequently, the mast cells were exposed to either 2.5 μM pigment or an equivalent amount of Δ cylE extract or 5 μM A23187 for 1 hour. Release of β-hex was then quantified in the mast cell supernatants. Data shown were obtained from three independent experiments performed in duplicate and compared to the respective untreated mast cells ( n = 3; ** P = 0.002, Dunnett’s multiple comparison test following ANOVA; error bars, ±SEM).
    Ca2 Indicator Fluo 4, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 537 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore ca2 ionophore a23187
    Mast cell degranulation by the GBS pigment requires <t>Ca2</t> + influx ( A ) PCMCs were pretreated with the calcium indicator Fluo-4-AM, and calcium influx was recorded by flow cytometry. At 60 s, either 5 μM A23187 (top panel), 0.5 μM GBS pigment (bottom panel), or an equivalent amount of control Δ cylE extract (middle panel) was added. Mean fluorescence intensities of mast cells before treatment (red) and after treatment (green) are shown. Time is given in seconds. A representative image from one of three independent experiments is shown. ( B ) PCMCs were pretreated with either EGTA (4 mM) or LY294002 (100 μM) for 30 min or with pertussis toxin (PT; 200 ng/ml) for 2 hours. Untreated PCMCs were included as controls for both pretreatment conditions. Subsequently, the mast cells were exposed to either 2.5 μM pigment or an equivalent amount of Δ cylE extract or 5 μM A23187 for 1 hour. Release of β-hex was then quantified in the mast cell supernatants. Data shown were obtained from three independent experiments performed in duplicate and compared to the respective untreated mast cells ( n = 3; ** P = 0.002, Dunnett’s multiple comparison test following ANOVA; error bars, ±SEM).
    Ca2 Ionophore A23187, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 301 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher ca2 indicator fura 2
    Mast cell degranulation by the GBS pigment requires <t>Ca2</t> + influx ( A ) PCMCs were pretreated with the calcium indicator Fluo-4-AM, and calcium influx was recorded by flow cytometry. At 60 s, either 5 μM A23187 (top panel), 0.5 μM GBS pigment (bottom panel), or an equivalent amount of control Δ cylE extract (middle panel) was added. Mean fluorescence intensities of mast cells before treatment (red) and after treatment (green) are shown. Time is given in seconds. A representative image from one of three independent experiments is shown. ( B ) PCMCs were pretreated with either EGTA (4 mM) or LY294002 (100 μM) for 30 min or with pertussis toxin (PT; 200 ng/ml) for 2 hours. Untreated PCMCs were included as controls for both pretreatment conditions. Subsequently, the mast cells were exposed to either 2.5 μM pigment or an equivalent amount of Δ cylE extract or 5 μM A23187 for 1 hour. Release of β-hex was then quantified in the mast cell supernatants. Data shown were obtained from three independent experiments performed in duplicate and compared to the respective untreated mast cells ( n = 3; ** P = 0.002, Dunnett’s multiple comparison test following ANOVA; error bars, ±SEM).
    Ca2 Indicator Fura 2, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 194 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Ipsen Group ca2 channel syntaxin interaction
    Mast cell degranulation by the GBS pigment requires <t>Ca2</t> + influx ( A ) PCMCs were pretreated with the calcium indicator Fluo-4-AM, and calcium influx was recorded by flow cytometry. At 60 s, either 5 μM A23187 (top panel), 0.5 μM GBS pigment (bottom panel), or an equivalent amount of control Δ cylE extract (middle panel) was added. Mean fluorescence intensities of mast cells before treatment (red) and after treatment (green) are shown. Time is given in seconds. A representative image from one of three independent experiments is shown. ( B ) PCMCs were pretreated with either EGTA (4 mM) or LY294002 (100 μM) for 30 min or with pertussis toxin (PT; 200 ng/ml) for 2 hours. Untreated PCMCs were included as controls for both pretreatment conditions. Subsequently, the mast cells were exposed to either 2.5 μM pigment or an equivalent amount of Δ cylE extract or 5 μM A23187 for 1 hour. Release of β-hex was then quantified in the mast cell supernatants. Data shown were obtained from three independent experiments performed in duplicate and compared to the respective untreated mast cells ( n = 3; ** P = 0.002, Dunnett’s multiple comparison test following ANOVA; error bars, ±SEM).
    Ca2 Channel Syntaxin Interaction, supplied by Ipsen Group, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Ipsen Group n type ca2
    Mast cell degranulation by the GBS pigment requires <t>Ca2</t> + influx ( A ) PCMCs were pretreated with the calcium indicator Fluo-4-AM, and calcium influx was recorded by flow cytometry. At 60 s, either 5 μM A23187 (top panel), 0.5 μM GBS pigment (bottom panel), or an equivalent amount of control Δ cylE extract (middle panel) was added. Mean fluorescence intensities of mast cells before treatment (red) and after treatment (green) are shown. Time is given in seconds. A representative image from one of three independent experiments is shown. ( B ) PCMCs were pretreated with either EGTA (4 mM) or LY294002 (100 μM) for 30 min or with pertussis toxin (PT; 200 ng/ml) for 2 hours. Untreated PCMCs were included as controls for both pretreatment conditions. Subsequently, the mast cells were exposed to either 2.5 μM pigment or an equivalent amount of Δ cylE extract or 5 μM A23187 for 1 hour. Release of β-hex was then quantified in the mast cell supernatants. Data shown were obtained from three independent experiments performed in duplicate and compared to the respective untreated mast cells ( n = 3; ** P = 0.002, Dunnett’s multiple comparison test following ANOVA; error bars, ±SEM).
    N Type Ca2, supplied by Ipsen Group, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher ca2 indicator fluo 3 am
    Mast cell degranulation by the GBS pigment requires <t>Ca2</t> + influx ( A ) PCMCs were pretreated with the calcium indicator Fluo-4-AM, and calcium influx was recorded by flow cytometry. At 60 s, either 5 μM A23187 (top panel), 0.5 μM GBS pigment (bottom panel), or an equivalent amount of control Δ cylE extract (middle panel) was added. Mean fluorescence intensities of mast cells before treatment (red) and after treatment (green) are shown. Time is given in seconds. A representative image from one of three independent experiments is shown. ( B ) PCMCs were pretreated with either EGTA (4 mM) or LY294002 (100 μM) for 30 min or with pertussis toxin (PT; 200 ng/ml) for 2 hours. Untreated PCMCs were included as controls for both pretreatment conditions. Subsequently, the mast cells were exposed to either 2.5 μM pigment or an equivalent amount of Δ cylE extract or 5 μM A23187 for 1 hour. Release of β-hex was then quantified in the mast cell supernatants. Data shown were obtained from three independent experiments performed in duplicate and compared to the respective untreated mast cells ( n = 3; ** P = 0.002, Dunnett’s multiple comparison test following ANOVA; error bars, ±SEM).
    Ca2 Indicator Fluo 3 Am, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Swant ca2 exchanger
    Mast cell degranulation by the GBS pigment requires <t>Ca2</t> + influx ( A ) PCMCs were pretreated with the calcium indicator Fluo-4-AM, and calcium influx was recorded by flow cytometry. At 60 s, either 5 μM A23187 (top panel), 0.5 μM GBS pigment (bottom panel), or an equivalent amount of control Δ cylE extract (middle panel) was added. Mean fluorescence intensities of mast cells before treatment (red) and after treatment (green) are shown. Time is given in seconds. A representative image from one of three independent experiments is shown. ( B ) PCMCs were pretreated with either EGTA (4 mM) or LY294002 (100 μM) for 30 min or with pertussis toxin (PT; 200 ng/ml) for 2 hours. Untreated PCMCs were included as controls for both pretreatment conditions. Subsequently, the mast cells were exposed to either 2.5 μM pigment or an equivalent amount of Δ cylE extract or 5 μM A23187 for 1 hour. Release of β-hex was then quantified in the mast cell supernatants. Data shown were obtained from three independent experiments performed in duplicate and compared to the respective untreated mast cells ( n = 3; ** P = 0.002, Dunnett’s multiple comparison test following ANOVA; error bars, ±SEM).
    Ca2 Exchanger, supplied by Swant, used in various techniques. Bioz Stars score: 88/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Ipsen Group p q type ca2
    Mast cell degranulation by the GBS pigment requires <t>Ca2</t> + influx ( A ) PCMCs were pretreated with the calcium indicator Fluo-4-AM, and calcium influx was recorded by flow cytometry. At 60 s, either 5 μM A23187 (top panel), 0.5 μM GBS pigment (bottom panel), or an equivalent amount of control Δ cylE extract (middle panel) was added. Mean fluorescence intensities of mast cells before treatment (red) and after treatment (green) are shown. Time is given in seconds. A representative image from one of three independent experiments is shown. ( B ) PCMCs were pretreated with either EGTA (4 mM) or LY294002 (100 μM) for 30 min or with pertussis toxin (PT; 200 ng/ml) for 2 hours. Untreated PCMCs were included as controls for both pretreatment conditions. Subsequently, the mast cells were exposed to either 2.5 μM pigment or an equivalent amount of Δ cylE extract or 5 μM A23187 for 1 hour. Release of β-hex was then quantified in the mast cell supernatants. Data shown were obtained from three independent experiments performed in duplicate and compared to the respective untreated mast cells ( n = 3; ** P = 0.002, Dunnett’s multiple comparison test following ANOVA; error bars, ±SEM).
    P Q Type Ca2, supplied by Ipsen Group, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Journal of Biological Chemistry ca2
    Mast cell degranulation by the GBS pigment requires <t>Ca2</t> + influx ( A ) PCMCs were pretreated with the calcium indicator Fluo-4-AM, and calcium influx was recorded by flow cytometry. At 60 s, either 5 μM A23187 (top panel), 0.5 μM GBS pigment (bottom panel), or an equivalent amount of control Δ cylE extract (middle panel) was added. Mean fluorescence intensities of mast cells before treatment (red) and after treatment (green) are shown. Time is given in seconds. A representative image from one of three independent experiments is shown. ( B ) PCMCs were pretreated with either EGTA (4 mM) or LY294002 (100 μM) for 30 min or with pertussis toxin (PT; 200 ng/ml) for 2 hours. Untreated PCMCs were included as controls for both pretreatment conditions. Subsequently, the mast cells were exposed to either 2.5 μM pigment or an equivalent amount of Δ cylE extract or 5 μM A23187 for 1 hour. Release of β-hex was then quantified in the mast cell supernatants. Data shown were obtained from three independent experiments performed in duplicate and compared to the respective untreated mast cells ( n = 3; ** P = 0.002, Dunnett’s multiple comparison test following ANOVA; error bars, ±SEM).
    Ca2, supplied by Journal of Biological Chemistry, used in various techniques. Bioz Stars score: 92/100, based on 53 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher ca2 indicator indo 1
    Mast cell degranulation by the GBS pigment requires <t>Ca2</t> + influx ( A ) PCMCs were pretreated with the calcium indicator Fluo-4-AM, and calcium influx was recorded by flow cytometry. At 60 s, either 5 μM A23187 (top panel), 0.5 μM GBS pigment (bottom panel), or an equivalent amount of control Δ cylE extract (middle panel) was added. Mean fluorescence intensities of mast cells before treatment (red) and after treatment (green) are shown. Time is given in seconds. A representative image from one of three independent experiments is shown. ( B ) PCMCs were pretreated with either EGTA (4 mM) or LY294002 (100 μM) for 30 min or with pertussis toxin (PT; 200 ng/ml) for 2 hours. Untreated PCMCs were included as controls for both pretreatment conditions. Subsequently, the mast cells were exposed to either 2.5 μM pigment or an equivalent amount of Δ cylE extract or 5 μM A23187 for 1 hour. Release of β-hex was then quantified in the mast cell supernatants. Data shown were obtained from three independent experiments performed in duplicate and compared to the respective untreated mast cells ( n = 3; ** P = 0.002, Dunnett’s multiple comparison test following ANOVA; error bars, ±SEM).
    Ca2 Indicator Indo 1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 106 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Ipsen Group ca2 influx
    Mast cell degranulation by the GBS pigment requires <t>Ca2</t> + influx ( A ) PCMCs were pretreated with the calcium indicator Fluo-4-AM, and calcium influx was recorded by flow cytometry. At 60 s, either 5 μM A23187 (top panel), 0.5 μM GBS pigment (bottom panel), or an equivalent amount of control Δ cylE extract (middle panel) was added. Mean fluorescence intensities of mast cells before treatment (red) and after treatment (green) are shown. Time is given in seconds. A representative image from one of three independent experiments is shown. ( B ) PCMCs were pretreated with either EGTA (4 mM) or LY294002 (100 μM) for 30 min or with pertussis toxin (PT; 200 ng/ml) for 2 hours. Untreated PCMCs were included as controls for both pretreatment conditions. Subsequently, the mast cells were exposed to either 2.5 μM pigment or an equivalent amount of Δ cylE extract or 5 μM A23187 for 1 hour. Release of β-hex was then quantified in the mast cell supernatants. Data shown were obtained from three independent experiments performed in duplicate and compared to the respective untreated mast cells ( n = 3; ** P = 0.002, Dunnett’s multiple comparison test following ANOVA; error bars, ±SEM).
    Ca2 Influx, supplied by Ipsen Group, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Medicago intranuclear ca2
    Mast cell degranulation by the GBS pigment requires <t>Ca2</t> + influx ( A ) PCMCs were pretreated with the calcium indicator Fluo-4-AM, and calcium influx was recorded by flow cytometry. At 60 s, either 5 μM A23187 (top panel), 0.5 μM GBS pigment (bottom panel), or an equivalent amount of control Δ cylE extract (middle panel) was added. Mean fluorescence intensities of mast cells before treatment (red) and after treatment (green) are shown. Time is given in seconds. A representative image from one of three independent experiments is shown. ( B ) PCMCs were pretreated with either EGTA (4 mM) or LY294002 (100 μM) for 30 min or with pertussis toxin (PT; 200 ng/ml) for 2 hours. Untreated PCMCs were included as controls for both pretreatment conditions. Subsequently, the mast cells were exposed to either 2.5 μM pigment or an equivalent amount of Δ cylE extract or 5 μM A23187 for 1 hour. Release of β-hex was then quantified in the mast cell supernatants. Data shown were obtained from three independent experiments performed in duplicate and compared to the respective untreated mast cells ( n = 3; ** P = 0.002, Dunnett’s multiple comparison test following ANOVA; error bars, ±SEM).
    Intranuclear Ca2, supplied by Medicago, used in various techniques. Bioz Stars score: 88/100, based on 62 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    N/A
    RayBio Custom Human CA2 ELISA Kit 5 7 week lead time
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    RayBio Human Immunoquantitative PCR Based CA2 ELISA Kit for cell culture supernatants plasma and serum samples
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    Carbonic anhydrases CAs are a large family of zinc metalloenzymes CAs form a family of enzymes that catalyze the rapid interconversion of carbon dioxide and water to bicarbonate and protons
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    Image Search Results


    Representative images and quantification of western blot analysis of calcium- and voltage-gated potassium channels. (A) Total expression of BK and SK2 potassium channels in mouse hippocampus showed no significant differences between WT and Cav2.3-KO mice (WT = 8, KO = 6; p > 0.05). In contrast, in the Cav2.3-KO, the total level of Kv4.2 was significantly increased compared to the WT (WT = 8, KO = 6; p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Functional Coupling of Cav2.3 and BK Potassium Channels Regulates Action Potential Repolarization and Short-Term Plasticity in the Mouse Hippocampus

    doi: 10.3389/fncel.2019.00027

    Figure Lengend Snippet: Representative images and quantification of western blot analysis of calcium- and voltage-gated potassium channels. (A) Total expression of BK and SK2 potassium channels in mouse hippocampus showed no significant differences between WT and Cav2.3-KO mice (WT = 8, KO = 6; p > 0.05). In contrast, in the Cav2.3-KO, the total level of Kv4.2 was significantly increased compared to the WT (WT = 8, KO = 6; p

    Article Snippet: The separated proteins were immuonoblotted using Cav2.3 (1:1,000, Synaptic Systems, Germany) or BK antibody (1:2,000, Alomone Labs, Israel) and visualized by Alexa Fluor 680 secondary antibody (1:10,000, Invitrogen, Carlsbad, CA, USA) and Alexa Fluor 800 secondary antibody (1:5,000, Rockland, Knox County, MA, USA).

    Techniques: Western Blot, Expressing, Mouse Assay

    AP waveform properties from CA1 pyramidal neurons. (A) Average first AP elicited by a +150 pA current injection for WT (black traces) and Cav2.3 KO (red traces) ± SEM. (B–G) Quantification of (A) . Firing threshold = first derivative of voltage > 0.2, and AP onset = time between current step onset and firing threshold. After-hyperpolarization (AHP) is calculated relative to firing threshold. (H) Example traces showing the first six APs elicited by a +200 pA current injection for WT (black traces) and Cav2.3 KO (red traces). (I–N) Quantification of (H) . Instead of AP onset, this analysis shows the inter-spike interval (time between peak voltages) and AHP was calculated as the minimum voltage between two APs. n = 17 for WT, and 19 for KO, all data shown as mean ± SEM, with Student’s T -Test to probe for significance and Holm-Sidak to account for multiple comparisons (n.s. = not significant, * p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Functional Coupling of Cav2.3 and BK Potassium Channels Regulates Action Potential Repolarization and Short-Term Plasticity in the Mouse Hippocampus

    doi: 10.3389/fncel.2019.00027

    Figure Lengend Snippet: AP waveform properties from CA1 pyramidal neurons. (A) Average first AP elicited by a +150 pA current injection for WT (black traces) and Cav2.3 KO (red traces) ± SEM. (B–G) Quantification of (A) . Firing threshold = first derivative of voltage > 0.2, and AP onset = time between current step onset and firing threshold. After-hyperpolarization (AHP) is calculated relative to firing threshold. (H) Example traces showing the first six APs elicited by a +200 pA current injection for WT (black traces) and Cav2.3 KO (red traces). (I–N) Quantification of (H) . Instead of AP onset, this analysis shows the inter-spike interval (time between peak voltages) and AHP was calculated as the minimum voltage between two APs. n = 17 for WT, and 19 for KO, all data shown as mean ± SEM, with Student’s T -Test to probe for significance and Holm-Sidak to account for multiple comparisons (n.s. = not significant, * p

    Article Snippet: The separated proteins were immuonoblotted using Cav2.3 (1:1,000, Synaptic Systems, Germany) or BK antibody (1:2,000, Alomone Labs, Israel) and visualized by Alexa Fluor 680 secondary antibody (1:10,000, Invitrogen, Carlsbad, CA, USA) and Alexa Fluor 800 secondary antibody (1:5,000, Rockland, Knox County, MA, USA).

    Techniques: Injection

    Altered synaptic connectivity between CA1 and subiculum. (A–D) Analysis of spontaneous excitatory postsynaptic currents (sEPSCs) recorded from subicular pyramidal neurons in the presence of Gabazine. (A) Representative traces from WT (black traces) and Cav2.3 KO (red traces) and cumulative probability of sEPSC amplitudes for all recorded events ( n > 5,000). Insets represent the average traces over all sEPSCs. (B) Quantification of amplitude and frequency for the average event recorded from each cell per genotype ( n = 20). (C) Cumulative distribution and quantification for the rise- and decay-times for the average sEPSCs. (D) Quantification for the charge conveyed with each sEPSC (area under the current trace). (E–H) Experimental setup (E) and analysis (F–H) of paired-pulse facilitation between CA1 and subiculum. (F,G) EPSP amplitude and decay time are not significantly different between WT and KO [analyzed from the first elicited EPSP of the 1,000 ms inter-stimulus intervals (ISIs) pulse]. Insets in (H) represent average traces for WT (black) and Cav2.3 KO (red) for two representative ISI. All data shown as mean ± SEM, with student’s T -Test or two-way ANOVA where appropriate, to probe for significance and Holm-Sidak to account for multiple comparisons (n.s. = not significant, * p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Functional Coupling of Cav2.3 and BK Potassium Channels Regulates Action Potential Repolarization and Short-Term Plasticity in the Mouse Hippocampus

    doi: 10.3389/fncel.2019.00027

    Figure Lengend Snippet: Altered synaptic connectivity between CA1 and subiculum. (A–D) Analysis of spontaneous excitatory postsynaptic currents (sEPSCs) recorded from subicular pyramidal neurons in the presence of Gabazine. (A) Representative traces from WT (black traces) and Cav2.3 KO (red traces) and cumulative probability of sEPSC amplitudes for all recorded events ( n > 5,000). Insets represent the average traces over all sEPSCs. (B) Quantification of amplitude and frequency for the average event recorded from each cell per genotype ( n = 20). (C) Cumulative distribution and quantification for the rise- and decay-times for the average sEPSCs. (D) Quantification for the charge conveyed with each sEPSC (area under the current trace). (E–H) Experimental setup (E) and analysis (F–H) of paired-pulse facilitation between CA1 and subiculum. (F,G) EPSP amplitude and decay time are not significantly different between WT and KO [analyzed from the first elicited EPSP of the 1,000 ms inter-stimulus intervals (ISIs) pulse]. Insets in (H) represent average traces for WT (black) and Cav2.3 KO (red) for two representative ISI. All data shown as mean ± SEM, with student’s T -Test or two-way ANOVA where appropriate, to probe for significance and Holm-Sidak to account for multiple comparisons (n.s. = not significant, * p

    Article Snippet: The separated proteins were immuonoblotted using Cav2.3 (1:1,000, Synaptic Systems, Germany) or BK antibody (1:2,000, Alomone Labs, Israel) and visualized by Alexa Fluor 680 secondary antibody (1:10,000, Invitrogen, Carlsbad, CA, USA) and Alexa Fluor 800 secondary antibody (1:5,000, Rockland, Knox County, MA, USA).

    Techniques: Mass Spectrometry

    Cav2.3 knockout (KO) CA1 pyramidal cells are hyperexcitable. (A) Resting membrane potential (Vrest; A1 ) and cell capacitance (A2) , measured after break-in. (B) Input resistance quantified as the slope of an IV-curve for hyperpolarizing current steps from −160 to −20 pA. Insets represent averaged voltage traces for wildtype (WT; black) and KO (red) recordings during current injection. (C) Hyperpolarization activated current I H , measured as sag-percentage from maximum voltage deflection (C1) and as the slope of the rebound potential after the end of a series of hyperpolarizing current injections plotted against the steady-state voltage during the current injection (C2) . (D) Firing frequency of action potentials (APs) in CA1 pyramidal neurons of Cav2.3 KO and WT animals, elicited by current steps from Vrest, by injection of positive or negative current in steps of 20 pA. Inset shows example recordings for injections of +200 pA WT (black traces) or Cav2.3 KO (red traces). n = 17 for WT, and 21 for KO. All data shown as mean ± SEM, with Student’s T -Test to probe for significance (n.s. = not significant, * p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Functional Coupling of Cav2.3 and BK Potassium Channels Regulates Action Potential Repolarization and Short-Term Plasticity in the Mouse Hippocampus

    doi: 10.3389/fncel.2019.00027

    Figure Lengend Snippet: Cav2.3 knockout (KO) CA1 pyramidal cells are hyperexcitable. (A) Resting membrane potential (Vrest; A1 ) and cell capacitance (A2) , measured after break-in. (B) Input resistance quantified as the slope of an IV-curve for hyperpolarizing current steps from −160 to −20 pA. Insets represent averaged voltage traces for wildtype (WT; black) and KO (red) recordings during current injection. (C) Hyperpolarization activated current I H , measured as sag-percentage from maximum voltage deflection (C1) and as the slope of the rebound potential after the end of a series of hyperpolarizing current injections plotted against the steady-state voltage during the current injection (C2) . (D) Firing frequency of action potentials (APs) in CA1 pyramidal neurons of Cav2.3 KO and WT animals, elicited by current steps from Vrest, by injection of positive or negative current in steps of 20 pA. Inset shows example recordings for injections of +200 pA WT (black traces) or Cav2.3 KO (red traces). n = 17 for WT, and 21 for KO. All data shown as mean ± SEM, with Student’s T -Test to probe for significance (n.s. = not significant, * p

    Article Snippet: The separated proteins were immuonoblotted using Cav2.3 (1:1,000, Synaptic Systems, Germany) or BK antibody (1:2,000, Alomone Labs, Israel) and visualized by Alexa Fluor 680 secondary antibody (1:10,000, Invitrogen, Carlsbad, CA, USA) and Alexa Fluor 800 secondary antibody (1:5,000, Rockland, Knox County, MA, USA).

    Techniques: Knock-Out, Injection

    Stimulus-induced secretion consists of CIVDS and CDS components in the somata of DRG neurons. ( A ) Depolarization- and UV-flash-induced C m signals in a DRG neuron dialyzed with 5 mM NP-EGTA and 0.2 mM fura-6F. In Ca2+-free extracellular solution ( left panel ), a step depolarization (200 ms) evoked CIVDS ( C m0 ). When extracellular Ca 2+ was added ( right panel ), UV flashes ( arrows ) evoked a substantial increase of C m ( C m (UV)), which was caused by the Ca 2+ increase in the absence of membrane depolarization. Series conductance ( G s) and membrane current ( I m ), and Ca 2+ trace ([Ca 2+ ]i) are also shown. ( B ) Depolarization-induced C m signals. In Ca 2+ -free extracellular solution ( left panel ), a step depolarization (200 ms) evoked a C m increase (CIVDS) followed by a rapid reversal as in A . C m1 refers to the entire course. Subsequently, extracellular Ca 2+ (2.5 mM) was puffed ( right panel ), and the same stimulus evoked a biphasic response ( C m2 ). The first Cm increase was followed by a small but rapid reversal and then a slow but substantial increase. C m2 − C m1 ( dashed line ) indicated the CDS component.

    Journal: Biophysical Journal

    Article Title: Action Potential Modulates Ca2+-Dependent and Ca2+-Independent Secretion in a Sensory Neuron

    doi: 10.1016/j.bpj.2008.11.037

    Figure Lengend Snippet: Stimulus-induced secretion consists of CIVDS and CDS components in the somata of DRG neurons. ( A ) Depolarization- and UV-flash-induced C m signals in a DRG neuron dialyzed with 5 mM NP-EGTA and 0.2 mM fura-6F. In Ca2+-free extracellular solution ( left panel ), a step depolarization (200 ms) evoked CIVDS ( C m0 ). When extracellular Ca 2+ was added ( right panel ), UV flashes ( arrows ) evoked a substantial increase of C m ( C m (UV)), which was caused by the Ca 2+ increase in the absence of membrane depolarization. Series conductance ( G s) and membrane current ( I m ), and Ca 2+ trace ([Ca 2+ ]i) are also shown. ( B ) Depolarization-induced C m signals. In Ca 2+ -free extracellular solution ( left panel ), a step depolarization (200 ms) evoked a C m increase (CIVDS) followed by a rapid reversal as in A . C m1 refers to the entire course. Subsequently, extracellular Ca 2+ (2.5 mM) was puffed ( right panel ), and the same stimulus evoked a biphasic response ( C m2 ). The first Cm increase was followed by a small but rapid reversal and then a slow but substantial increase. C m2 − C m1 ( dashed line ) indicated the CDS component.

    Article Snippet: Alteration of Ca2+ dependence of neurotransmitter release by disruption of Ca2+ channel/syntaxin interaction.

    Techniques: Mass Spectrometry

    Mast cell degranulation by the GBS pigment requires Ca2 + influx ( A ) PCMCs were pretreated with the calcium indicator Fluo-4-AM, and calcium influx was recorded by flow cytometry. At 60 s, either 5 μM A23187 (top panel), 0.5 μM GBS pigment (bottom panel), or an equivalent amount of control Δ cylE extract (middle panel) was added. Mean fluorescence intensities of mast cells before treatment (red) and after treatment (green) are shown. Time is given in seconds. A representative image from one of three independent experiments is shown. ( B ) PCMCs were pretreated with either EGTA (4 mM) or LY294002 (100 μM) for 30 min or with pertussis toxin (PT; 200 ng/ml) for 2 hours. Untreated PCMCs were included as controls for both pretreatment conditions. Subsequently, the mast cells were exposed to either 2.5 μM pigment or an equivalent amount of Δ cylE extract or 5 μM A23187 for 1 hour. Release of β-hex was then quantified in the mast cell supernatants. Data shown were obtained from three independent experiments performed in duplicate and compared to the respective untreated mast cells ( n = 3; ** P = 0.002, Dunnett’s multiple comparison test following ANOVA; error bars, ±SEM).

    Journal: Science advances

    Article Title: Mast cell degranulation by a hemolytic lipid toxin decreases GBS colonization and infection

    doi: 10.1126/sciadv.1400225

    Figure Lengend Snippet: Mast cell degranulation by the GBS pigment requires Ca2 + influx ( A ) PCMCs were pretreated with the calcium indicator Fluo-4-AM, and calcium influx was recorded by flow cytometry. At 60 s, either 5 μM A23187 (top panel), 0.5 μM GBS pigment (bottom panel), or an equivalent amount of control Δ cylE extract (middle panel) was added. Mean fluorescence intensities of mast cells before treatment (red) and after treatment (green) are shown. Time is given in seconds. A representative image from one of three independent experiments is shown. ( B ) PCMCs were pretreated with either EGTA (4 mM) or LY294002 (100 μM) for 30 min or with pertussis toxin (PT; 200 ng/ml) for 2 hours. Untreated PCMCs were included as controls for both pretreatment conditions. Subsequently, the mast cells were exposed to either 2.5 μM pigment or an equivalent amount of Δ cylE extract or 5 μM A23187 for 1 hour. Release of β-hex was then quantified in the mast cell supernatants. Data shown were obtained from three independent experiments performed in duplicate and compared to the respective untreated mast cells ( n = 3; ** P = 0.002, Dunnett’s multiple comparison test following ANOVA; error bars, ±SEM).

    Article Snippet: Briefly, 2 × 105 PCMCs were loaded with 5 μM of the fluorescent Ca2+ indicator (Fluo-4-AM, Life Technologies) for 30 min, and cells were then washed and resuspended in Tyrode’s buffer without BSA.

    Techniques: Flow Cytometry, Cytometry, Fluorescence