paxilline  (Alomone Labs)


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    Structured Review

    Alomone Labs paxilline
    Effects of Ca 2+ chelators and Ca 2+ channel inhibitors on BK currents recorded from SCN neurons during the day and night. <t>Paxilline-sensitive</t> macroscopic BK currents were recorded from C57BL6 WT (A–E) and Ca V 1.3 WT (F) SCNs. All intracellular solutions in this study were made with 0.9 mM EGTA, except where 5 mM BAPTA was substituted (A–E) . Currents were elicited from a holding potential of −90 mV by 150-ms voltage steps from −110 to +90 mV in +20-mV increments. (A,B) Representative BK currents from −90 to +90 mV are shown from day (A) and night (B) SCN neurons. (C,D) Current-voltage plot comparing BK current density recorded in either control EGTA or BAPTA during the day (C) and night (D) . (E) Summary of BK current density at +90 mV recorded with control EGTA or BAPTA with Ca 2+ channel inhibitors 10 μM nimodipine (Nim) during the day, or 10 μM dantrolene (Dan) at night. In EGTA, Nim decreased BK currents compared to controls ( P = 0.0008), with no significant difference in BAPTA. At night, BK currents were decreased with BAPTA compared to control EGTA conditions ( P
    Paxilline, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Contributions of CaV1.3 Channels to Ca2+ Current and Ca2+-Activated BK Current in the Suprachiasmatic Nucleus"

    Article Title: Contributions of CaV1.3 Channels to Ca2+ Current and Ca2+-Activated BK Current in the Suprachiasmatic Nucleus

    Journal: Frontiers in Physiology

    doi: 10.3389/fphys.2021.737291

    Effects of Ca 2+ chelators and Ca 2+ channel inhibitors on BK currents recorded from SCN neurons during the day and night. Paxilline-sensitive macroscopic BK currents were recorded from C57BL6 WT (A–E) and Ca V 1.3 WT (F) SCNs. All intracellular solutions in this study were made with 0.9 mM EGTA, except where 5 mM BAPTA was substituted (A–E) . Currents were elicited from a holding potential of −90 mV by 150-ms voltage steps from −110 to +90 mV in +20-mV increments. (A,B) Representative BK currents from −90 to +90 mV are shown from day (A) and night (B) SCN neurons. (C,D) Current-voltage plot comparing BK current density recorded in either control EGTA or BAPTA during the day (C) and night (D) . (E) Summary of BK current density at +90 mV recorded with control EGTA or BAPTA with Ca 2+ channel inhibitors 10 μM nimodipine (Nim) during the day, or 10 μM dantrolene (Dan) at night. In EGTA, Nim decreased BK currents compared to controls ( P = 0.0008), with no significant difference in BAPTA. At night, BK currents were decreased with BAPTA compared to control EGTA conditions ( P
    Figure Legend Snippet: Effects of Ca 2+ chelators and Ca 2+ channel inhibitors on BK currents recorded from SCN neurons during the day and night. Paxilline-sensitive macroscopic BK currents were recorded from C57BL6 WT (A–E) and Ca V 1.3 WT (F) SCNs. All intracellular solutions in this study were made with 0.9 mM EGTA, except where 5 mM BAPTA was substituted (A–E) . Currents were elicited from a holding potential of −90 mV by 150-ms voltage steps from −110 to +90 mV in +20-mV increments. (A,B) Representative BK currents from −90 to +90 mV are shown from day (A) and night (B) SCN neurons. (C,D) Current-voltage plot comparing BK current density recorded in either control EGTA or BAPTA during the day (C) and night (D) . (E) Summary of BK current density at +90 mV recorded with control EGTA or BAPTA with Ca 2+ channel inhibitors 10 μM nimodipine (Nim) during the day, or 10 μM dantrolene (Dan) at night. In EGTA, Nim decreased BK currents compared to controls ( P = 0.0008), with no significant difference in BAPTA. At night, BK currents were decreased with BAPTA compared to control EGTA conditions ( P

    Techniques Used:

    BK currents from Ca V 1.3 WT and Ca V 1.3 KO SCN during the day and night. Paxilline-sensitive macroscopic BK currents were recorded as in Figure 1 . (A) Summary of BK current densities at +90 mV recorded from Ca V 1.3 WT and Ca V 1.3 KO neurons during the day and night. BK currents were increased at night for both Ca V 1.3 WT ( P = 0.004) and Ca V 1.3 KO ( P = 0.007) compared to the day. * P
    Figure Legend Snippet: BK currents from Ca V 1.3 WT and Ca V 1.3 KO SCN during the day and night. Paxilline-sensitive macroscopic BK currents were recorded as in Figure 1 . (A) Summary of BK current densities at +90 mV recorded from Ca V 1.3 WT and Ca V 1.3 KO neurons during the day and night. BK currents were increased at night for both Ca V 1.3 WT ( P = 0.004) and Ca V 1.3 KO ( P = 0.007) compared to the day. * P

    Techniques Used:

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    Alomone Labs mouse anti bk channel α
    Membrane current in  HEK  293 cells stably expressing human  BK  channel α‐ and β1‐subunits.  (A)  Voltage‐dependent  BK  current traces recorded in a representative cell with the voltage protocol as shown in the  inset  before and after application of 1 μM paxilline (a selective  BK  channel blocker), and washout of paxilline.  (B)  I‐V  relationships of  BK  current in the absence and presence of 1 μM paxilline, and upon washout ( n  = 5, * P 
    Mouse Anti Bk Channel α, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs anti bk β4
    Effects of <t>BK-β4</t> small interfering (si) RNA on shear-stress (10 dynes/cm 2 )-induced K efflux from C11. A : C11 were untreated (normal), transfected with green fluorescent protein (control), 4 nontarget siRNA oligos (siRNA nontarget), or 6 siRNA
    Anti Bk β4, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs bk channel α subunits
    Functional expression of BK channels in rat MV. A, co-expression of BK channel α- and β1-subunits in rat MV. Myocytes labeled with an anti-BK channel <t>α-subunits</t> (anti- slo ) antibody (green) and anti-BKCa 2+ β1 (red). B , Western
    Bk Channel α Subunits, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs bk β1 subunit
    Mitochondrial calcium uptake is impaired by Paxilline and by genetic ablation of <t>β1-subunit</t> A) Paxilline reduces the amount of mitochondrial Ca 2+ necessary to induce formation and opening of mPTP. B) Mitochondria from the β1 KO required less Ca 2+ than mitochondria from the Wt to trigger the opening of mPTP. C) Calcium retention capacity values obtained in A for paxilline and in B for the β1-KO mitochondria. Bars SEM, (* p
    Bk β1 Subunit, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Membrane current in  HEK  293 cells stably expressing human  BK  channel α‐ and β1‐subunits.  (A)  Voltage‐dependent  BK  current traces recorded in a representative cell with the voltage protocol as shown in the  inset  before and after application of 1 μM paxilline (a selective  BK  channel blocker), and washout of paxilline.  (B)  I‐V  relationships of  BK  current in the absence and presence of 1 μM paxilline, and upon washout ( n  = 5, * P 

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Tyrphostin AG556 increases the activity of large conductance Ca2+‐activated K+ channels by inhibiting epidermal growth factor receptor tyrosine kinase

    doi: 10.1111/jcmm.13103

    Figure Lengend Snippet: Membrane current in HEK 293 cells stably expressing human BK channel α‐ and β1‐subunits. (A) Voltage‐dependent BK current traces recorded in a representative cell with the voltage protocol as shown in the inset before and after application of 1 μM paxilline (a selective BK channel blocker), and washout of paxilline. (B) I‐V relationships of BK current in the absence and presence of 1 μM paxilline, and upon washout ( n  = 5, * P 

    Article Snippet: Proteins were immunoprecipitated overnight at 4°C using 1 μg of mouse anti‐BK channel α (APC‐021; Alomone Labs, Jerusalem, Israel) antibody or 1 μg of mouse anti‐β1 (APC‐036; Alomone Labs, Jerusalem, Israel) antibody and 20 μl of Protein A/G beads (sc‐2003; Santa Cruz Biotechnology, Inc. CA, USA).

    Techniques: Stable Transfection, Expressing

    Effects of BK-β4 small interfering (si) RNA on shear-stress (10 dynes/cm 2 )-induced K efflux from C11. A : C11 were untreated (normal), transfected with green fluorescent protein (control), 4 nontarget siRNA oligos (siRNA nontarget), or 6 siRNA

    Journal: American Journal of Physiology - Renal Physiology

    Article Title: Shear stress-induced volume decrease in C11-MDCK cells by BK-?/?4

    doi: 10.1152/ajprenal.00222.2010

    Figure Lengend Snippet: Effects of BK-β4 small interfering (si) RNA on shear-stress (10 dynes/cm 2 )-induced K efflux from C11. A : C11 were untreated (normal), transfected with green fluorescent protein (control), 4 nontarget siRNA oligos (siRNA nontarget), or 6 siRNA

    Article Snippet: Primary antibodies included anti-BK-α (rabbit polyclonal, diluted 1:1,000, Alomone Labs), anti-BK-β4 (rabbit polyclonal, diluted 1:1,000, Alomone Labs), and anti-actin (rabbit polyclonal, 1:1,000, Santa Cruz Biotechnology, Santa Cruz, CA) with either donkey (Santa Cruz Biotechnology) or goat (Pierce Biotechnology) anti-rabbit IgG-conjugated horseradish peroxidase secondary antibody diluted 1:20,000–1:40,000.

    Techniques: Transfection

    Functional expression of BK channels in rat MV. A, co-expression of BK channel α- and β1-subunits in rat MV. Myocytes labeled with an anti-BK channel α-subunits (anti- slo ) antibody (green) and anti-BKCa 2+ β1 (red). B , Western

    Journal: Journal of cardiovascular pharmacology

    Article Title: Requirement for functional BK channels in maintaining oscillation in venomotor tone revealed by species differences in expression of the ?1 accessory subunits

    doi: 10.1097/FJC.0b013e318233614c

    Figure Lengend Snippet: Functional expression of BK channels in rat MV. A, co-expression of BK channel α- and β1-subunits in rat MV. Myocytes labeled with an anti-BK channel α-subunits (anti- slo ) antibody (green) and anti-BKCa 2+ β1 (red). B , Western

    Article Snippet: For Western blot analysis, we used a primary antibody targeted to BK channel α-subunits (anti-BKca2+ 1.1, 1:500; Alomone Labs.

    Techniques: Functional Assay, Expressing, Labeling, Western Blot

    Mitochondrial calcium uptake is impaired by Paxilline and by genetic ablation of β1-subunit A) Paxilline reduces the amount of mitochondrial Ca 2+ necessary to induce formation and opening of mPTP. B) Mitochondria from the β1 KO required less Ca 2+ than mitochondria from the Wt to trigger the opening of mPTP. C) Calcium retention capacity values obtained in A for paxilline and in B for the β1-KO mitochondria. Bars SEM, (* p

    Journal: The Journal of physiology

    Article Title: MitoBKCa channel is functionally associated with its regulatory β1 subunit in cardiac mitochondria

    doi: 10.1113/JP277769

    Figure Lengend Snippet: Mitochondrial calcium uptake is impaired by Paxilline and by genetic ablation of β1-subunit A) Paxilline reduces the amount of mitochondrial Ca 2+ necessary to induce formation and opening of mPTP. B) Mitochondria from the β1 KO required less Ca 2+ than mitochondria from the Wt to trigger the opening of mPTP. C) Calcium retention capacity values obtained in A for paxilline and in B for the β1-KO mitochondria. Bars SEM, (* p

    Article Snippet: Polyclonal antibody against BKCa (Rabbit Anti-KCNMA1 Cat. APC-021 and APC-151), and polyclonal antibody for BK-β1 subunit (Rabbit Anti-KCNMB1, Cat. APC-036) were from Alomone labs.

    Techniques:

    MitoBK Ca alpha and auxiliary β1 subunit association in cardiac mitochondria. A) Immunoprecipitation (IP) of BK Ca alpha and BK-β1 subunit with an antibody against BK Ca ). Taken together, these results indicates an association between mitoBK Ca and its auxiliary β1 subunit. Having demonstrated this, we characterized the biophysical properties of mitoBK Ca in Wt and β1-KO cardiac mitoplasts. B) Single-channel currents of mitoBK Ca recorded from Wt mitoplasts. The P o of the channel was 0.9 ± 0.08, n=4, in average at +80 mV and the indicated matrix Ca 2+ . C) Representative traces of a mitoplast from the β1 KO, were no single-channel current was detected at the indicated voltages and matrix Ca 2+ (upper traces). Few mitoplasts (5 out of 28, mitoplasts, n=5 hearts) showed mitoBK Ca channel activity with a P o of 0.4 ± 0.004, n=3 at +40 mV in the indicated matrix Ca 2+ . D) Plots of I vs. V at 12 µM matrix Ca 2+ from Wt and β1 KO mitoplasts (data from 6 and 3 patches, respectively). A slope conductance of 302±26 (n=4) and 335 ±15 pS (n=3) was calculated for Wt and β1 KO, respectively. E) Plots of P o vs. V for mitoBK Ca channels, data points represent the average of 4 and 3 mitoplasts for Wt and BK-β1 KO, respectively. Bars SEM.

    Journal: The Journal of physiology

    Article Title: MitoBKCa channel is functionally associated with its regulatory β1 subunit in cardiac mitochondria

    doi: 10.1113/JP277769

    Figure Lengend Snippet: MitoBK Ca alpha and auxiliary β1 subunit association in cardiac mitochondria. A) Immunoprecipitation (IP) of BK Ca alpha and BK-β1 subunit with an antibody against BK Ca ). Taken together, these results indicates an association between mitoBK Ca and its auxiliary β1 subunit. Having demonstrated this, we characterized the biophysical properties of mitoBK Ca in Wt and β1-KO cardiac mitoplasts. B) Single-channel currents of mitoBK Ca recorded from Wt mitoplasts. The P o of the channel was 0.9 ± 0.08, n=4, in average at +80 mV and the indicated matrix Ca 2+ . C) Representative traces of a mitoplast from the β1 KO, were no single-channel current was detected at the indicated voltages and matrix Ca 2+ (upper traces). Few mitoplasts (5 out of 28, mitoplasts, n=5 hearts) showed mitoBK Ca channel activity with a P o of 0.4 ± 0.004, n=3 at +40 mV in the indicated matrix Ca 2+ . D) Plots of I vs. V at 12 µM matrix Ca 2+ from Wt and β1 KO mitoplasts (data from 6 and 3 patches, respectively). A slope conductance of 302±26 (n=4) and 335 ±15 pS (n=3) was calculated for Wt and β1 KO, respectively. E) Plots of P o vs. V for mitoBK Ca channels, data points represent the average of 4 and 3 mitoplasts for Wt and BK-β1 KO, respectively. Bars SEM.

    Article Snippet: Polyclonal antibody against BKCa (Rabbit Anti-KCNMA1 Cat. APC-021 and APC-151), and polyclonal antibody for BK-β1 subunit (Rabbit Anti-KCNMB1, Cat. APC-036) were from Alomone labs.

    Techniques: Immunoprecipitation, Activity Assay

    Localization of BKDEC and β1 subunit in HeLa cells. A) Cultured cells co-transfected with BKDEC alpha+β1subunit, loaded with mitotracker red, and DAPI B) β1-flag labeled with an anti-flag C) BKDEC labeled with anti-HA. D) Merge of mitotracker red, BKDEC and β1. E) Cross-correlation index (CCi), relative to mitotracker red signal, calculated from cells overexpressing BKDEC alone (Cci=0.53±0.05) and co-expressed with β1 subunit (Cci=0.77±0.01, n=11 cells from 3 different preparations).*p

    Journal: The Journal of physiology

    Article Title: MitoBKCa channel is functionally associated with its regulatory β1 subunit in cardiac mitochondria

    doi: 10.1113/JP277769

    Figure Lengend Snippet: Localization of BKDEC and β1 subunit in HeLa cells. A) Cultured cells co-transfected with BKDEC alpha+β1subunit, loaded with mitotracker red, and DAPI B) β1-flag labeled with an anti-flag C) BKDEC labeled with anti-HA. D) Merge of mitotracker red, BKDEC and β1. E) Cross-correlation index (CCi), relative to mitotracker red signal, calculated from cells overexpressing BKDEC alone (Cci=0.53±0.05) and co-expressed with β1 subunit (Cci=0.77±0.01, n=11 cells from 3 different preparations).*p

    Article Snippet: Polyclonal antibody against BKCa (Rabbit Anti-KCNMA1 Cat. APC-021 and APC-151), and polyclonal antibody for BK-β1 subunit (Rabbit Anti-KCNMB1, Cat. APC-036) were from Alomone labs.

    Techniques: Cell Culture, Transfection, Labeling