k252a  (Alomone Labs)


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    Structured Review

    Alomone Labs k252a
    Mechanisms of NGF action. A , Example traces showing mEPSCs from cholinergic neurons grown in control (left) versus NGF condition (right). B , Kolmogorov–Smirnov graphs illustrate that NGF decreased the interevent interval (left) without altering the amplitude (right) of mEPSCs in cholinergic neurons ( n = 2 control, 3 NGF neurons). C , D , MLR2/3 (p75 NTR -antibodies), but not <t>K252a</t> (trk inhibitor), prevented NGF-induced increase in the amplitude of EPSCs in cholinergic neurons. n.s., p > 0.05. * p
    K252a, 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
    https://www.bioz.com/result/k252a/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
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    Images

    1) Product Images from "Chronic Exposure to Nerve Growth Factor Increases Acetylcholine and Glutamate Release from Cholinergic Neurons of the Rat Medial Septum and Diagonal Band of Broca via Mechanisms Mediated by p75NTR"

    Article Title: Chronic Exposure to Nerve Growth Factor Increases Acetylcholine and Glutamate Release from Cholinergic Neurons of the Rat Medial Septum and Diagonal Band of Broca via Mechanisms Mediated by p75NTR

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.4851-07.2008

    Mechanisms of NGF action. A , Example traces showing mEPSCs from cholinergic neurons grown in control (left) versus NGF condition (right). B , Kolmogorov–Smirnov graphs illustrate that NGF decreased the interevent interval (left) without altering the amplitude (right) of mEPSCs in cholinergic neurons ( n = 2 control, 3 NGF neurons). C , D , MLR2/3 (p75 NTR -antibodies), but not K252a (trk inhibitor), prevented NGF-induced increase in the amplitude of EPSCs in cholinergic neurons. n.s., p > 0.05. * p
    Figure Legend Snippet: Mechanisms of NGF action. A , Example traces showing mEPSCs from cholinergic neurons grown in control (left) versus NGF condition (right). B , Kolmogorov–Smirnov graphs illustrate that NGF decreased the interevent interval (left) without altering the amplitude (right) of mEPSCs in cholinergic neurons ( n = 2 control, 3 NGF neurons). C , D , MLR2/3 (p75 NTR -antibodies), but not K252a (trk inhibitor), prevented NGF-induced increase in the amplitude of EPSCs in cholinergic neurons. n.s., p > 0.05. * p

    Techniques Used:

    NGF increased both acetylcholine and glutamate transmission from cholinergic neurons via p75 NTR . A , Cholinergic neurons exposed to NGF displayed larger nicotinic and glutamatergic currents than controls. B , C , K252a did not affect NGF-induced increases in nicotinic or glutamatergic currents. D , E , MLR2/3 significantly reduced the effects of NGF on nicotinic and glutamatergic currents. F , G , Cholinergic neurons grown in NGF had significantly larger cell bodies compared with controls. The effect of NGF on soma size was blocked by K252a, but not by MLR2/3. n.s., p > 0.05. * p
    Figure Legend Snippet: NGF increased both acetylcholine and glutamate transmission from cholinergic neurons via p75 NTR . A , Cholinergic neurons exposed to NGF displayed larger nicotinic and glutamatergic currents than controls. B , C , K252a did not affect NGF-induced increases in nicotinic or glutamatergic currents. D , E , MLR2/3 significantly reduced the effects of NGF on nicotinic and glutamatergic currents. F , G , Cholinergic neurons grown in NGF had significantly larger cell bodies compared with controls. The effect of NGF on soma size was blocked by K252a, but not by MLR2/3. n.s., p > 0.05. * p

    Techniques Used: Transmission Assay

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    Alomone Labs anti kv1 5
    Characterization of the Kv1.3 and <t>Kv1.5</t> mutant channels containing the YS segment. A , average normalized activation and inactivation curves are shown as conductance-voltage relationships for Kv1.3, Kv1.5, the truncated Kv1.3-YS channel, and the chimeras Kv1.5-YS 532 and Kv1.5-YS 613 . All datasets were fitted to Boltzmann functions. Each data point is the mean ± S.E. of 6–11 cells. B , confocal images of non-permeabilized cells transfected with Kv1.3-YS-Cherry, Kv1.5-YS 532 -EGFP, and Kv1.5-YS 613 -EGFP. An extracellular anti-Kv1.3 antibody was used to label Kv1.3-YS ( green ), whereas the extracellular anti-Kv1.5 antibody was used for Kv1.5-YS 532 and Kv1.5-YS 613 chimeras ( red ). Nuclei were stained by Hoechst ( blue ). C , proliferation rate of the indicated channels or GFP-transfected cells (control) was determined by measuring EdU incorporation. Significant differences when comparing to Kv1.3 (*) or to control (#) are indicated. Statistical analysis was performed with one-way ANOVA followed by a Tukey's HSD multiple comparison. Each bar is the average of 9–15 determinations from 5 different assays. D , the average peak current amplitude obtained in cell-attached experiments for Kv1.5 channels and all the Kv1.5 chimeras was plotted against the % of the channels expressed at the plasma membrane ( upper graph ) or their normalized effect on proliferation (taking 100% as the proliferation rate of GFP-transfected HEK cells, lower graph ). The correlation between expression and current was fit to a linear regression curve ( y = 18.54 + 0.0066x, R 2 = 0.85, p = 0.008), but there was no correlation between proliferation and current amplitude ( R 2 = 0.23, p = 0.19).
    Anti Kv1 5, 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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    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti kv1 5 - by Bioz Stars, 2022-07
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    Characterization of the Kv1.3 and Kv1.5 mutant channels containing the YS segment. A , average normalized activation and inactivation curves are shown as conductance-voltage relationships for Kv1.3, Kv1.5, the truncated Kv1.3-YS channel, and the chimeras Kv1.5-YS 532 and Kv1.5-YS 613 . All datasets were fitted to Boltzmann functions. Each data point is the mean ± S.E. of 6–11 cells. B , confocal images of non-permeabilized cells transfected with Kv1.3-YS-Cherry, Kv1.5-YS 532 -EGFP, and Kv1.5-YS 613 -EGFP. An extracellular anti-Kv1.3 antibody was used to label Kv1.3-YS ( green ), whereas the extracellular anti-Kv1.5 antibody was used for Kv1.5-YS 532 and Kv1.5-YS 613 chimeras ( red ). Nuclei were stained by Hoechst ( blue ). C , proliferation rate of the indicated channels or GFP-transfected cells (control) was determined by measuring EdU incorporation. Significant differences when comparing to Kv1.3 (*) or to control (#) are indicated. Statistical analysis was performed with one-way ANOVA followed by a Tukey's HSD multiple comparison. Each bar is the average of 9–15 determinations from 5 different assays. D , the average peak current amplitude obtained in cell-attached experiments for Kv1.5 channels and all the Kv1.5 chimeras was plotted against the % of the channels expressed at the plasma membrane ( upper graph ) or their normalized effect on proliferation (taking 100% as the proliferation rate of GFP-transfected HEK cells, lower graph ). The correlation between expression and current was fit to a linear regression curve ( y = 18.54 + 0.0066x, R 2 = 0.85, p = 0.008), but there was no correlation between proliferation and current amplitude ( R 2 = 0.23, p = 0.19).

    Journal: The Journal of Biological Chemistry

    Article Title: Molecular Determinants of Kv1.3 Potassium Channels-induced Proliferation *

    doi: 10.1074/jbc.M115.678995

    Figure Lengend Snippet: Characterization of the Kv1.3 and Kv1.5 mutant channels containing the YS segment. A , average normalized activation and inactivation curves are shown as conductance-voltage relationships for Kv1.3, Kv1.5, the truncated Kv1.3-YS channel, and the chimeras Kv1.5-YS 532 and Kv1.5-YS 613 . All datasets were fitted to Boltzmann functions. Each data point is the mean ± S.E. of 6–11 cells. B , confocal images of non-permeabilized cells transfected with Kv1.3-YS-Cherry, Kv1.5-YS 532 -EGFP, and Kv1.5-YS 613 -EGFP. An extracellular anti-Kv1.3 antibody was used to label Kv1.3-YS ( green ), whereas the extracellular anti-Kv1.5 antibody was used for Kv1.5-YS 532 and Kv1.5-YS 613 chimeras ( red ). Nuclei were stained by Hoechst ( blue ). C , proliferation rate of the indicated channels or GFP-transfected cells (control) was determined by measuring EdU incorporation. Significant differences when comparing to Kv1.3 (*) or to control (#) are indicated. Statistical analysis was performed with one-way ANOVA followed by a Tukey's HSD multiple comparison. Each bar is the average of 9–15 determinations from 5 different assays. D , the average peak current amplitude obtained in cell-attached experiments for Kv1.5 channels and all the Kv1.5 chimeras was plotted against the % of the channels expressed at the plasma membrane ( upper graph ) or their normalized effect on proliferation (taking 100% as the proliferation rate of GFP-transfected HEK cells, lower graph ). The correlation between expression and current was fit to a linear regression curve ( y = 18.54 + 0.0066x, R 2 = 0.85, p = 0.008), but there was no correlation between proliferation and current amplitude ( R 2 = 0.23, p = 0.19).

    Article Snippet: Non-permeabilized cells were incubated with anti-Kv1.3 or anti Kv1.5 extracellular primary antibodies (APC101 or APC150, Alomone Labs), whereas permeabilized cells were incubated with anti-Kv1.3 COOH (75-009, NeuroMab) or anti-Kv1.5 COOH (APC004, Alomone Labs), all at a final concentration of 1:50.

    Techniques: Mutagenesis, Activation Assay, Transfection, Staining, Expressing

    Mutant KCNA5 is located in perinuclear packets and not on the cell surface of transfected HEK-293 cells and human (h)PASMC. A : HEK-293 cells transfected with WT KCNA5 ( a and c ) or G182R ( b and d ) were stained with anti-KCNA5 antibody (Ab-KCNA5, red) and

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Tetramerization domain mutations in KCNA5 affect channel kinetics and cause abnormal trafficking patterns

    doi: 10.1152/ajpcell.00464.2009

    Figure Lengend Snippet: Mutant KCNA5 is located in perinuclear packets and not on the cell surface of transfected HEK-293 cells and human (h)PASMC. A : HEK-293 cells transfected with WT KCNA5 ( a and c ) or G182R ( b and d ) were stained with anti-KCNA5 antibody (Ab-KCNA5, red) and

    Article Snippet: After cells were fixed in 4% paraformaldehyde-PBS for 15 min at room temperature, coverslips were incubated in blocking solution (2% BSA, 2% FBS, 0.1% Triton X-100 in PBS) for 1 h. Coverslips were then exposed to rabbit anti-KV1.5 antibody (Alomone Labs) at a dilution of 1:100 in blocking solution for 2 h at room temperature.

    Techniques: Mutagenesis, Transfection, Staining

    G182R and E211D mutations cause incomplete processing of KCNA5 in HEK-293 cells. HEK-293 cells were transfected with WT KCNA5, G182R, E211D, or G182R/E211D and subjected to standard immunoblot procedures. A : representative immunoblot from cells transfected

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Tetramerization domain mutations in KCNA5 affect channel kinetics and cause abnormal trafficking patterns

    doi: 10.1152/ajpcell.00464.2009

    Figure Lengend Snippet: G182R and E211D mutations cause incomplete processing of KCNA5 in HEK-293 cells. HEK-293 cells were transfected with WT KCNA5, G182R, E211D, or G182R/E211D and subjected to standard immunoblot procedures. A : representative immunoblot from cells transfected

    Article Snippet: After cells were fixed in 4% paraformaldehyde-PBS for 15 min at room temperature, coverslips were incubated in blocking solution (2% BSA, 2% FBS, 0.1% Triton X-100 in PBS) for 1 h. Coverslips were then exposed to rabbit anti-KV1.5 antibody (Alomone Labs) at a dilution of 1:100 in blocking solution for 2 h at room temperature.

    Techniques: Transfection

    Mutant KCNA5 forms functional homotetrameric channels. A : COS-1, HEK-293, and human pulmonary artery smooth muscle cells (PASMC) were transiently transfected with either empty vector [green fluorescent protein (GFP)] or WT KCNA5 (WT) as indicated. Whole

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Tetramerization domain mutations in KCNA5 affect channel kinetics and cause abnormal trafficking patterns

    doi: 10.1152/ajpcell.00464.2009

    Figure Lengend Snippet: Mutant KCNA5 forms functional homotetrameric channels. A : COS-1, HEK-293, and human pulmonary artery smooth muscle cells (PASMC) were transiently transfected with either empty vector [green fluorescent protein (GFP)] or WT KCNA5 (WT) as indicated. Whole

    Article Snippet: After cells were fixed in 4% paraformaldehyde-PBS for 15 min at room temperature, coverslips were incubated in blocking solution (2% BSA, 2% FBS, 0.1% Triton X-100 in PBS) for 1 h. Coverslips were then exposed to rabbit anti-KV1.5 antibody (Alomone Labs) at a dilution of 1:100 in blocking solution for 2 h at room temperature.

    Techniques: Mutagenesis, Functional Assay, Transfection, Plasmid Preparation

    Mutations in KCNA5 at G182 and E211 do not affect the pharmacological effect of 4-aminopyridine (4-AP). A : a standard I-V pulse protocol was delivered to HEK-293 cells transiently transfected with the indicated vector [WT KCNA5 ( a ), G182R ( b ), E211D (

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Tetramerization domain mutations in KCNA5 affect channel kinetics and cause abnormal trafficking patterns

    doi: 10.1152/ajpcell.00464.2009

    Figure Lengend Snippet: Mutations in KCNA5 at G182 and E211 do not affect the pharmacological effect of 4-aminopyridine (4-AP). A : a standard I-V pulse protocol was delivered to HEK-293 cells transiently transfected with the indicated vector [WT KCNA5 ( a ), G182R ( b ), E211D (

    Article Snippet: After cells were fixed in 4% paraformaldehyde-PBS for 15 min at room temperature, coverslips were incubated in blocking solution (2% BSA, 2% FBS, 0.1% Triton X-100 in PBS) for 1 h. Coverslips were then exposed to rabbit anti-KV1.5 antibody (Alomone Labs) at a dilution of 1:100 in blocking solution for 2 h at room temperature.

    Techniques: Transfection, Plasmid Preparation

    Decreased G182R expression in COS-1 cells cannot be rescued by overexpression of K V β subunits. A , a : HEK-293 cells were transfected with WT KCNA5 or K V β1.3-HA alone or cotransfected with WT KCNA5, G182R, E211D, or G182R/E211D and K V β1.3-HA.

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Tetramerization domain mutations in KCNA5 affect channel kinetics and cause abnormal trafficking patterns

    doi: 10.1152/ajpcell.00464.2009

    Figure Lengend Snippet: Decreased G182R expression in COS-1 cells cannot be rescued by overexpression of K V β subunits. A , a : HEK-293 cells were transfected with WT KCNA5 or K V β1.3-HA alone or cotransfected with WT KCNA5, G182R, E211D, or G182R/E211D and K V β1.3-HA.

    Article Snippet: After cells were fixed in 4% paraformaldehyde-PBS for 15 min at room temperature, coverslips were incubated in blocking solution (2% BSA, 2% FBS, 0.1% Triton X-100 in PBS) for 1 h. Coverslips were then exposed to rabbit anti-KV1.5 antibody (Alomone Labs) at a dilution of 1:100 in blocking solution for 2 h at room temperature.

    Techniques: Expressing, Over Expression, Transfection

    Two nonsynonymous mutations identified in the KCNA5 gene from idiopathic pulmonary arterial hypertension (IPAH) patients localize to the NH 2 -terminal tetramerization domain (T1 domain). A , left : schematic diagram of voltage-gated K + (K V ) channel subunit

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Tetramerization domain mutations in KCNA5 affect channel kinetics and cause abnormal trafficking patterns

    doi: 10.1152/ajpcell.00464.2009

    Figure Lengend Snippet: Two nonsynonymous mutations identified in the KCNA5 gene from idiopathic pulmonary arterial hypertension (IPAH) patients localize to the NH 2 -terminal tetramerization domain (T1 domain). A , left : schematic diagram of voltage-gated K + (K V ) channel subunit

    Article Snippet: After cells were fixed in 4% paraformaldehyde-PBS for 15 min at room temperature, coverslips were incubated in blocking solution (2% BSA, 2% FBS, 0.1% Triton X-100 in PBS) for 1 h. Coverslips were then exposed to rabbit anti-KV1.5 antibody (Alomone Labs) at a dilution of 1:100 in blocking solution for 2 h at room temperature.

    Techniques:

    Cotransfection of K V β subunits affects KCNA5 channel kinetics. HEK-293 cells were transfected with WT KCNA5 alone (KCNA5) or in the presence of K V β1.3-hemagglutinin (HA) (KCNA5/K V β1.3). A and B : representative current recordings

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Tetramerization domain mutations in KCNA5 affect channel kinetics and cause abnormal trafficking patterns

    doi: 10.1152/ajpcell.00464.2009

    Figure Lengend Snippet: Cotransfection of K V β subunits affects KCNA5 channel kinetics. HEK-293 cells were transfected with WT KCNA5 alone (KCNA5) or in the presence of K V β1.3-hemagglutinin (HA) (KCNA5/K V β1.3). A and B : representative current recordings

    Article Snippet: After cells were fixed in 4% paraformaldehyde-PBS for 15 min at room temperature, coverslips were incubated in blocking solution (2% BSA, 2% FBS, 0.1% Triton X-100 in PBS) for 1 h. Coverslips were then exposed to rabbit anti-KV1.5 antibody (Alomone Labs) at a dilution of 1:100 in blocking solution for 2 h at room temperature.

    Techniques: Cotransfection, Transfection

    Voltage-dependent inactivation is accelerated in the G182R mutant KCNA5 channel. A standard 2-pulse inactivation protocol was used to determine channel availability after a 10-s prepulse in HEK-293 cells transiently transfected with WT KCNA5, G182R, E211D,

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Tetramerization domain mutations in KCNA5 affect channel kinetics and cause abnormal trafficking patterns

    doi: 10.1152/ajpcell.00464.2009

    Figure Lengend Snippet: Voltage-dependent inactivation is accelerated in the G182R mutant KCNA5 channel. A standard 2-pulse inactivation protocol was used to determine channel availability after a 10-s prepulse in HEK-293 cells transiently transfected with WT KCNA5, G182R, E211D,

    Article Snippet: After cells were fixed in 4% paraformaldehyde-PBS for 15 min at room temperature, coverslips were incubated in blocking solution (2% BSA, 2% FBS, 0.1% Triton X-100 in PBS) for 1 h. Coverslips were then exposed to rabbit anti-KV1.5 antibody (Alomone Labs) at a dilution of 1:100 in blocking solution for 2 h at room temperature.

    Techniques: Mutagenesis, Transfection

    G182R protein expression is significantly decreased in COS-1 cells. A : COS-1 cells were transiently transfected with water (Mock), WT-KCNA5, G182R, E211D, or G182R/E211D. Representative images are shown at ×40 magnification. B : transfected cells

    Journal: American Journal of Physiology - Cell Physiology

    Article Title: Tetramerization domain mutations in KCNA5 affect channel kinetics and cause abnormal trafficking patterns

    doi: 10.1152/ajpcell.00464.2009

    Figure Lengend Snippet: G182R protein expression is significantly decreased in COS-1 cells. A : COS-1 cells were transiently transfected with water (Mock), WT-KCNA5, G182R, E211D, or G182R/E211D. Representative images are shown at ×40 magnification. B : transfected cells

    Article Snippet: After cells were fixed in 4% paraformaldehyde-PBS for 15 min at room temperature, coverslips were incubated in blocking solution (2% BSA, 2% FBS, 0.1% Triton X-100 in PBS) for 1 h. Coverslips were then exposed to rabbit anti-KV1.5 antibody (Alomone Labs) at a dilution of 1:100 in blocking solution for 2 h at room temperature.

    Techniques: Expressing, Transfection

    Kv3.4 in the axonal growth cones of dorsal spinal commissural neurons. A–F , Transverse sections of the spinal cord of chick embryos were immunostained for Kv3.4. A , Absence of Kv3.4-IR in the dorsal spinal cord at HH17. Kv3.4-IR in precrossing commissural axons ( B–F , arrowheads) is evident during HH19-HH25 but disappears at HH27. D , Arrows indicate postcrossing commissural axons projecting from the other side of spinal cord. FP, Floor plate. G–L , Transverse sections of the spinal cord at HH23 were immunostained as indicated. G , Absence of Kv1.5-IR. H , Kv4.2-IR in the somata and dendrites of motoneurons (MN). I , Kv4.3-IR in the bifurcation zone (BZ). In addition to the BZ, Kv3.1b-IR is strong in postcrossing commissural axons ( J , arrow) but weak in precrossing commissural axons ( J , arrowhead). K , Absence of Kv3.2-IR. L , Kv3.3 in motoneurons. M–M″ , Double staining in transverse sections of the spinal cord at HH21 shows colocalization of Kv3.4 and axonin-1 in the growth cones (arrowheads) of commissural axons. N–N″ , Colocalization of Kv3.4 and axonin-1 in cultured dorsal spinal neurons isolated from HH21-HH23 chick embryos. O–P″ , Red fluorescence-tagged phalloidin colabeling reveals enrichment of Kv3.4 in the growth cone ( O–O″ ) and Kv3.1b in the soma/axon shaft ( P–P″ ) of cultured dorsal spinal neurons. Q–Q″ , Kv3.4 and DiI colabeling. White represents Kv3.4-abundant regions. Blue represents Kv3.4-sparse regions ( Q″ ). R , Ratio of Kv3.4/DiI in the soma, axon shaft, or growth cone of each neuron was obtained by dividing the fluorescence intensity of Kv3.4 by that of DiI. Data are mean ± SEM ( n = 8 neurons, pooled from three independent experiments done on different days). *** p

    Journal: The Journal of Neuroscience

    Article Title: K+ Channel Kv3.4 Is Essential for Axon Growth by Limiting the Influx of Ca2+ into Growth Cones

    doi: 10.1523/JNEUROSCI.1076-16.2017

    Figure Lengend Snippet: Kv3.4 in the axonal growth cones of dorsal spinal commissural neurons. A–F , Transverse sections of the spinal cord of chick embryos were immunostained for Kv3.4. A , Absence of Kv3.4-IR in the dorsal spinal cord at HH17. Kv3.4-IR in precrossing commissural axons ( B–F , arrowheads) is evident during HH19-HH25 but disappears at HH27. D , Arrows indicate postcrossing commissural axons projecting from the other side of spinal cord. FP, Floor plate. G–L , Transverse sections of the spinal cord at HH23 were immunostained as indicated. G , Absence of Kv1.5-IR. H , Kv4.2-IR in the somata and dendrites of motoneurons (MN). I , Kv4.3-IR in the bifurcation zone (BZ). In addition to the BZ, Kv3.1b-IR is strong in postcrossing commissural axons ( J , arrow) but weak in precrossing commissural axons ( J , arrowhead). K , Absence of Kv3.2-IR. L , Kv3.3 in motoneurons. M–M″ , Double staining in transverse sections of the spinal cord at HH21 shows colocalization of Kv3.4 and axonin-1 in the growth cones (arrowheads) of commissural axons. N–N″ , Colocalization of Kv3.4 and axonin-1 in cultured dorsal spinal neurons isolated from HH21-HH23 chick embryos. O–P″ , Red fluorescence-tagged phalloidin colabeling reveals enrichment of Kv3.4 in the growth cone ( O–O″ ) and Kv3.1b in the soma/axon shaft ( P–P″ ) of cultured dorsal spinal neurons. Q–Q″ , Kv3.4 and DiI colabeling. White represents Kv3.4-abundant regions. Blue represents Kv3.4-sparse regions ( Q″ ). R , Ratio of Kv3.4/DiI in the soma, axon shaft, or growth cone of each neuron was obtained by dividing the fluorescence intensity of Kv3.4 by that of DiI. Data are mean ± SEM ( n = 8 neurons, pooled from three independent experiments done on different days). *** p

    Article Snippet: After wash in low-salt Tris-buffered saline (LTBS; 25 m m Tris, 0.85% NaCl, pH 7.5) and then LTBS containing 0.1% Triton X-100 (LTBST), sections on slides were treated with 0.2% hydrogen peroxide in LTBS for 30 min. Nonspecific binding was blocked by 3% normal donkey serum and 2% BSA in LTBST for 1.5 h. Primary antibodies included rabbit anti-Kv1.5 (1:100), rabbit anti-Kv3.1b (1:100), rabbit anti-Kv3.2 (1:100), rabbit anti-Kv3.3 (1:100), rabbit anti-Kv3.4 (1:100), rabbit anti-Kv4.2 (1:250), and rabbit anti-Kv4.3 (1:1500).

    Techniques: Double Staining, Cell Culture, Isolation, Fluorescence

    Some Kv1.5 channel subunits are localized in lipid raft fractions in adult rat atrial myocytes Western blot analysis of the distribution of Kv1.5 subunits, connexin-43 and caveolin-3 on step sucrose gradient of proteins from atrial myocardium.

    Journal: The Journal of Physiology

    Article Title: Membrane cholesterol modulates Kv1.5 potassium channel distribution and function in rat cardiomyocytes

    doi: 10.1113/jphysiol.2007.134809

    Figure Lengend Snippet: Some Kv1.5 channel subunits are localized in lipid raft fractions in adult rat atrial myocytes Western blot analysis of the distribution of Kv1.5 subunits, connexin-43 and caveolin-3 on step sucrose gradient of proteins from atrial myocardium.

    Article Snippet: Then slides were incubated with rabbit anti-Kv1.5 polyclonal antibodies (pAbs) (1: 20), with mouse anti-caveolin-3 monoclonal antibodies (mAbs) (1: 50), with a mixture of mouse anti-caveolin-3 and rabbit anti-connexin-43 (1: 50) antibodies, or with a mixture of mouse anti-α-actinin sarcomeric and rabbit anti-Kv1.5 antibodies for 1 h at RT.

    Techniques: Western Blot

    Kv1.5 do not co-localized with caveolin-3 in adult atrial tissue Immunolocalization of Kv1.5 subunits ( A ) and caveolin-3 ( B ) in cryosections of atrial myocardium. Double immunostaining of connexin-43 (FITC, C ) and caveolin-3 (Texas Red, D ) in cryosections of atrial myocardium. E , merged image of the same area of the section in C and D , showing the lack of overlap between connexin-43 and caveolin-3 stainings.

    Journal: The Journal of Physiology

    Article Title: Membrane cholesterol modulates Kv1.5 potassium channel distribution and function in rat cardiomyocytes

    doi: 10.1113/jphysiol.2007.134809

    Figure Lengend Snippet: Kv1.5 do not co-localized with caveolin-3 in adult atrial tissue Immunolocalization of Kv1.5 subunits ( A ) and caveolin-3 ( B ) in cryosections of atrial myocardium. Double immunostaining of connexin-43 (FITC, C ) and caveolin-3 (Texas Red, D ) in cryosections of atrial myocardium. E , merged image of the same area of the section in C and D , showing the lack of overlap between connexin-43 and caveolin-3 stainings.

    Article Snippet: Then slides were incubated with rabbit anti-Kv1.5 polyclonal antibodies (pAbs) (1: 20), with mouse anti-caveolin-3 monoclonal antibodies (mAbs) (1: 50), with a mixture of mouse anti-caveolin-3 and rabbit anti-connexin-43 (1: 50) antibodies, or with a mixture of mouse anti-α-actinin sarcomeric and rabbit anti-Kv1.5 antibodies for 1 h at RT.

    Techniques: Double Immunostaining

    Surface expression of Kv1.5 subunits in neonatal cardiomyocytes A , in live cardiomyocytes, transfected GFP-tagged Kv1.5 subunits are clustered at the membrane surface adjacent to the bottom of laminin-coated glass support, as shown in the projection of Z sections in the lower panel. In contrast, GFP alone was homogeneously distributed in cardiomyocytes (inset). B , after the application of 2% MCD, clusters increased in size and were redistributed throughout the plasma membrane. C , bar graphs summarizing changes in cluster size upon MCD exposures; data are from 21 cardiomyocytes in control, and following incubation with 2% MCD for 7 min and 1 h 30 min. ** P

    Journal: The Journal of Physiology

    Article Title: Membrane cholesterol modulates Kv1.5 potassium channel distribution and function in rat cardiomyocytes

    doi: 10.1113/jphysiol.2007.134809

    Figure Lengend Snippet: Surface expression of Kv1.5 subunits in neonatal cardiomyocytes A , in live cardiomyocytes, transfected GFP-tagged Kv1.5 subunits are clustered at the membrane surface adjacent to the bottom of laminin-coated glass support, as shown in the projection of Z sections in the lower panel. In contrast, GFP alone was homogeneously distributed in cardiomyocytes (inset). B , after the application of 2% MCD, clusters increased in size and were redistributed throughout the plasma membrane. C , bar graphs summarizing changes in cluster size upon MCD exposures; data are from 21 cardiomyocytes in control, and following incubation with 2% MCD for 7 min and 1 h 30 min. ** P

    Article Snippet: Then slides were incubated with rabbit anti-Kv1.5 polyclonal antibodies (pAbs) (1: 20), with mouse anti-caveolin-3 monoclonal antibodies (mAbs) (1: 50), with a mixture of mouse anti-caveolin-3 and rabbit anti-connexin-43 (1: 50) antibodies, or with a mixture of mouse anti-α-actinin sarcomeric and rabbit anti-Kv1.5 antibodies for 1 h at RT.

    Techniques: Expressing, Transfection, Incubation

    Effect of cholesterol depletion on outward current parameters resulting from Kv1.5 subunit overexpression in neonatal cardiomyocytes Current density–voltage relationships ( A ) and voltage dependence ( B ) of I Kur activation under control conditions (•) and following 7 min MCD application (^). In A and B , each point represents average data from 5 cells.

    Journal: The Journal of Physiology

    Article Title: Membrane cholesterol modulates Kv1.5 potassium channel distribution and function in rat cardiomyocytes

    doi: 10.1113/jphysiol.2007.134809

    Figure Lengend Snippet: Effect of cholesterol depletion on outward current parameters resulting from Kv1.5 subunit overexpression in neonatal cardiomyocytes Current density–voltage relationships ( A ) and voltage dependence ( B ) of I Kur activation under control conditions (•) and following 7 min MCD application (^). In A and B , each point represents average data from 5 cells.

    Article Snippet: Then slides were incubated with rabbit anti-Kv1.5 polyclonal antibodies (pAbs) (1: 20), with mouse anti-caveolin-3 monoclonal antibodies (mAbs) (1: 50), with a mixture of mouse anti-caveolin-3 and rabbit anti-connexin-43 (1: 50) antibodies, or with a mixture of mouse anti-α-actinin sarcomeric and rabbit anti-Kv1.5 antibodies for 1 h at RT.

    Techniques: Over Expression, Activation Assay