rabbit anti kv7 2 antibody  (Alomone Labs)


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    Alomone Labs rabbit anti kv7 2 antibody
    NP1 interacts and colocalizes with <t>Kv7.2</t> at presynaptic terminals of excitatory synapses and axonal growth cones. A – D , Representative Western blots of immunoprecipitation eluates separated in 6% Tris-Glycine (for high molecular weight proteins) and 4–12% Bis-Tris gels (for low molecular weight proteins). TL, Total protein lysate; Syx, syntaxin. A , Both native Kv7.2 and syntaxin 1A, but not Kv7.3, coprecipitate with NP1 in total brain extracts. B , Both native Kv7.2 and NP1 coprecipitate with syntaxin in total brain extracts. C , D , Recombinant NP1 coprecipitates Kv7.2, but not syntaxin or Kv7.3, in 293T cells transfected with NP1, 5Myc-Kv7.2, 2HA-Kv7.3, and syntaxin 1A cDNAs. Kv7.2 and Kv7.3 were immunoprecipitated with antibodies against their respective Myc and HA tags. E , F , Immunofluorescence studies and confocal microscopy were performed in high-density ( E ) or low-density ( F ) isolated cortical neurons. E , Top, Confocal sections of 0.772 μm in the z -plane showing immunofluorescence of NP1, VGLUT1, Kv7.2, and negative control (omitting primary antibodies). Bottom, Colocalization (in white) of the excitatory presynaptic marker VGLUT1 (blue) with NP1 (green) and Kv7.2 (red) is shown in a single section with the corresponding orthogonal views of the stack of confocal sections. White arrows indicate sites of colocalization. F , NP1 (green) and KV7.2 (magenta) immunofluorescence and DIC images of an isolated cortical cultured neuron (1×) with its corresponding axonal growth cone highlighted in a white square box, shown in a confocal section of 0.772 μm in the z -plane at higher (5×) magnification. The negative control for primary antibodies is shown in another growth cone on the right. The image in the bottom is the merge of NP1 and Kv7.2 immunofluorescence images in the single confocal section obtained at 5× showing colocalization (white) of NP1 and Kv7.2 in the growth cone, with the corresponding orthogonal views from its respective stack of confocal sections. Images were acquired using restricted spectral emission wavelength ranges chosen to avoid crosstalk or bleed-through between the three different channels. Scale bar, 5 μm.
    Rabbit Anti Kv7 2 Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti kv7 2 antibody/product/Alomone Labs
    Average 93 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    rabbit anti kv7 2 antibody - by Bioz Stars, 2022-08
    93/100 stars

    Images

    1) Product Images from "Neuronal Pentraxin 1 Negatively Regulates Excitatory Synapse Density and Synaptic Plasticity"

    Article Title: Neuronal Pentraxin 1 Negatively Regulates Excitatory Synapse Density and Synaptic Plasticity

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.2548-14.2015

    NP1 interacts and colocalizes with Kv7.2 at presynaptic terminals of excitatory synapses and axonal growth cones. A – D , Representative Western blots of immunoprecipitation eluates separated in 6% Tris-Glycine (for high molecular weight proteins) and 4–12% Bis-Tris gels (for low molecular weight proteins). TL, Total protein lysate; Syx, syntaxin. A , Both native Kv7.2 and syntaxin 1A, but not Kv7.3, coprecipitate with NP1 in total brain extracts. B , Both native Kv7.2 and NP1 coprecipitate with syntaxin in total brain extracts. C , D , Recombinant NP1 coprecipitates Kv7.2, but not syntaxin or Kv7.3, in 293T cells transfected with NP1, 5Myc-Kv7.2, 2HA-Kv7.3, and syntaxin 1A cDNAs. Kv7.2 and Kv7.3 were immunoprecipitated with antibodies against their respective Myc and HA tags. E , F , Immunofluorescence studies and confocal microscopy were performed in high-density ( E ) or low-density ( F ) isolated cortical neurons. E , Top, Confocal sections of 0.772 μm in the z -plane showing immunofluorescence of NP1, VGLUT1, Kv7.2, and negative control (omitting primary antibodies). Bottom, Colocalization (in white) of the excitatory presynaptic marker VGLUT1 (blue) with NP1 (green) and Kv7.2 (red) is shown in a single section with the corresponding orthogonal views of the stack of confocal sections. White arrows indicate sites of colocalization. F , NP1 (green) and KV7.2 (magenta) immunofluorescence and DIC images of an isolated cortical cultured neuron (1×) with its corresponding axonal growth cone highlighted in a white square box, shown in a confocal section of 0.772 μm in the z -plane at higher (5×) magnification. The negative control for primary antibodies is shown in another growth cone on the right. The image in the bottom is the merge of NP1 and Kv7.2 immunofluorescence images in the single confocal section obtained at 5× showing colocalization (white) of NP1 and Kv7.2 in the growth cone, with the corresponding orthogonal views from its respective stack of confocal sections. Images were acquired using restricted spectral emission wavelength ranges chosen to avoid crosstalk or bleed-through between the three different channels. Scale bar, 5 μm.
    Figure Legend Snippet: NP1 interacts and colocalizes with Kv7.2 at presynaptic terminals of excitatory synapses and axonal growth cones. A – D , Representative Western blots of immunoprecipitation eluates separated in 6% Tris-Glycine (for high molecular weight proteins) and 4–12% Bis-Tris gels (for low molecular weight proteins). TL, Total protein lysate; Syx, syntaxin. A , Both native Kv7.2 and syntaxin 1A, but not Kv7.3, coprecipitate with NP1 in total brain extracts. B , Both native Kv7.2 and NP1 coprecipitate with syntaxin in total brain extracts. C , D , Recombinant NP1 coprecipitates Kv7.2, but not syntaxin or Kv7.3, in 293T cells transfected with NP1, 5Myc-Kv7.2, 2HA-Kv7.3, and syntaxin 1A cDNAs. Kv7.2 and Kv7.3 were immunoprecipitated with antibodies against their respective Myc and HA tags. E , F , Immunofluorescence studies and confocal microscopy were performed in high-density ( E ) or low-density ( F ) isolated cortical neurons. E , Top, Confocal sections of 0.772 μm in the z -plane showing immunofluorescence of NP1, VGLUT1, Kv7.2, and negative control (omitting primary antibodies). Bottom, Colocalization (in white) of the excitatory presynaptic marker VGLUT1 (blue) with NP1 (green) and Kv7.2 (red) is shown in a single section with the corresponding orthogonal views of the stack of confocal sections. White arrows indicate sites of colocalization. F , NP1 (green) and KV7.2 (magenta) immunofluorescence and DIC images of an isolated cortical cultured neuron (1×) with its corresponding axonal growth cone highlighted in a white square box, shown in a confocal section of 0.772 μm in the z -plane at higher (5×) magnification. The negative control for primary antibodies is shown in another growth cone on the right. The image in the bottom is the merge of NP1 and Kv7.2 immunofluorescence images in the single confocal section obtained at 5× showing colocalization (white) of NP1 and Kv7.2 in the growth cone, with the corresponding orthogonal views from its respective stack of confocal sections. Images were acquired using restricted spectral emission wavelength ranges chosen to avoid crosstalk or bleed-through between the three different channels. Scale bar, 5 μm.

    Techniques Used: Western Blot, Immunoprecipitation, Molecular Weight, Recombinant, Transfection, Immunofluorescence, Confocal Microscopy, Isolation, Negative Control, Marker, Cell Culture

    Knockdown of NP1 reduces Kv7.2 surface expression and M-current. A , B , The proportion of Kv7.2 at the neuronal surface was determined by biotinylation (30 min at 4°C) of all surface proteins in cultured cortical neurons. The ratio of the surface over the total amount of receptor was determined by densitometric analysis and expressed as a percentage of control values. Only samples of biotinylated proteins lacking immunoreactivity to actin, which indicates contamination with intracellular proteins, were used for analyses. A , Representative Western blots of Kv7.2, Na-KATPase, and actin from total (T) and biotinylated (B) proteins. B , Quantitative analysis of the effect of gain and loss of NP1 on the surface/total levels of the Kv7.2 subunit of the voltage-gated Kv7/M K + channels. Values are mean ± SEM of shRdm ( n = 11), shNP1 ( n = 11), and pWPI-NP1 ( n = 5), from at least three independent experiments; * p
    Figure Legend Snippet: Knockdown of NP1 reduces Kv7.2 surface expression and M-current. A , B , The proportion of Kv7.2 at the neuronal surface was determined by biotinylation (30 min at 4°C) of all surface proteins in cultured cortical neurons. The ratio of the surface over the total amount of receptor was determined by densitometric analysis and expressed as a percentage of control values. Only samples of biotinylated proteins lacking immunoreactivity to actin, which indicates contamination with intracellular proteins, were used for analyses. A , Representative Western blots of Kv7.2, Na-KATPase, and actin from total (T) and biotinylated (B) proteins. B , Quantitative analysis of the effect of gain and loss of NP1 on the surface/total levels of the Kv7.2 subunit of the voltage-gated Kv7/M K + channels. Values are mean ± SEM of shRdm ( n = 11), shNP1 ( n = 11), and pWPI-NP1 ( n = 5), from at least three independent experiments; * p

    Techniques Used: Expressing, Cell Culture, Western Blot

    2) Product Images from "Acute Stress Diminishes M-current Contributing to Elevated Activity of Hypothalamic-Pituitary-Adrenal Axis"

    Article Title: Acute Stress Diminishes M-current Contributing to Elevated Activity of Hypothalamic-Pituitary-Adrenal Axis

    Journal: Neuropharmacology

    doi: 10.1016/j.neuropharm.2016.11.024

    Inhibition of AMPK activity restored acute stress-induced decrease in membrane fraction of Kv7.3 subunit A and B: Gel images ( A ) and quantification of band density normalized by GAPDH ( B ) show the total protein levels of AMPK in the PVN in unstressed and stressed rats. C and D , Original gel images ( C ) and quantification ( D ) of GAPDH-normalized band density of membrane fraction of Kv7.2 and Kv7.3 in the PVN in stressed rats treated with ICV injection of vehicle and dorsomorphin (100 nmol in 10 µl aCSF). E and F , Original gel images ( E ) and quantification ( F ) of GAPDH-normalized band density of cytosolic fraction of Kv7.2 and Kv7.3 in the PVN in stressed rats treated with vehicle and dorsomorphin. The molecular weight was indicated on the right side of the gel images. In all the protein assays, 4 samples (each sample contains PVN tissues of 3 rats) were used in each group. *p
    Figure Legend Snippet: Inhibition of AMPK activity restored acute stress-induced decrease in membrane fraction of Kv7.3 subunit A and B: Gel images ( A ) and quantification of band density normalized by GAPDH ( B ) show the total protein levels of AMPK in the PVN in unstressed and stressed rats. C and D , Original gel images ( C ) and quantification ( D ) of GAPDH-normalized band density of membrane fraction of Kv7.2 and Kv7.3 in the PVN in stressed rats treated with ICV injection of vehicle and dorsomorphin (100 nmol in 10 µl aCSF). E and F , Original gel images ( E ) and quantification ( F ) of GAPDH-normalized band density of cytosolic fraction of Kv7.2 and Kv7.3 in the PVN in stressed rats treated with vehicle and dorsomorphin. The molecular weight was indicated on the right side of the gel images. In all the protein assays, 4 samples (each sample contains PVN tissues of 3 rats) were used in each group. *p

    Techniques Used: Inhibition, Activity Assay, Injection, Molecular Weight

    Acute stress diminished excitatory effect of M-current blocker and reduced M-current in PVN-CRH neurons ( A ): Confocal images show the distribution of Kv7.2 or Kv7.3 in the PVN-CRH neurons. B and C : Original recordings ( B ) and summary data ( C ) show the firing activity of eGFP-tagged PVN neurons in basal condition, the presence of Kv7 channel blocker XE-991 (3 µM), and washout in unstressed and acutely stressed rats (n = 20 neurons in unstressed and stressed rats, respectively). ( D) : Acute stress significantly reduced the increase in firing activity induced by XE-991. ( E) : Representative traces show deactivation tail currents in the absence (basal) or presence of Kv7 opener retigabine (10 µM) and blocker XE-991 (3 µM) in eGFP-tagged PVN neurons in stressed and unstressed rats. The deactivation tail currents were elicited by the voltage protocol shown in the insert. ( F ): summary data shows the basal and retigabine-induced M-currents, which were defined as XE-991-sensitive tail currents in basal and the presence of retigabine, were significantly diminished in the PVN-CRH neurons in stressed rats (n =15 neurons in 4 rats) compared with unstressed rats (n = 12 neurons in 4 rats).* p
    Figure Legend Snippet: Acute stress diminished excitatory effect of M-current blocker and reduced M-current in PVN-CRH neurons ( A ): Confocal images show the distribution of Kv7.2 or Kv7.3 in the PVN-CRH neurons. B and C : Original recordings ( B ) and summary data ( C ) show the firing activity of eGFP-tagged PVN neurons in basal condition, the presence of Kv7 channel blocker XE-991 (3 µM), and washout in unstressed and acutely stressed rats (n = 20 neurons in unstressed and stressed rats, respectively). ( D) : Acute stress significantly reduced the increase in firing activity induced by XE-991. ( E) : Representative traces show deactivation tail currents in the absence (basal) or presence of Kv7 opener retigabine (10 µM) and blocker XE-991 (3 µM) in eGFP-tagged PVN neurons in stressed and unstressed rats. The deactivation tail currents were elicited by the voltage protocol shown in the insert. ( F ): summary data shows the basal and retigabine-induced M-currents, which were defined as XE-991-sensitive tail currents in basal and the presence of retigabine, were significantly diminished in the PVN-CRH neurons in stressed rats (n =15 neurons in 4 rats) compared with unstressed rats (n = 12 neurons in 4 rats).* p

    Techniques Used: Activity Assay

    Kv7.2 and Kv7.3 protein expression levels in the PVN in stressed and unstressed rats A and B , Representative blots ( A ) and quantification ( B ) show the total protein levels of Kv7.2 and Kv7.3 in the PVN in unstressed and stressed rats. C and D , Original gel images ( C ) and quantification ( D ) show the membrane fraction protein levels of Kv7.2 and Kv7.3 in the PVN in unstressed and stressed rats. E and F , Original gel images ( E ) and quantification ( F ) show the cytosolic fraction protein levels of Kv7.2 and Kv7.3 in the PVN in unstressed and stressed rats (n = 4 samples, each sample contains PVN tissues of 3 rats) were used in each group. * p
    Figure Legend Snippet: Kv7.2 and Kv7.3 protein expression levels in the PVN in stressed and unstressed rats A and B , Representative blots ( A ) and quantification ( B ) show the total protein levels of Kv7.2 and Kv7.3 in the PVN in unstressed and stressed rats. C and D , Original gel images ( C ) and quantification ( D ) show the membrane fraction protein levels of Kv7.2 and Kv7.3 in the PVN in unstressed and stressed rats. E and F , Original gel images ( E ) and quantification ( F ) show the cytosolic fraction protein levels of Kv7.2 and Kv7.3 in the PVN in unstressed and stressed rats (n = 4 samples, each sample contains PVN tissues of 3 rats) were used in each group. * p

    Techniques Used: Expressing

    3) Product Images from "Neuronal Pentraxin 1 Negatively Regulates Excitatory Synapse Density and Synaptic Plasticity"

    Article Title: Neuronal Pentraxin 1 Negatively Regulates Excitatory Synapse Density and Synaptic Plasticity

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.2548-14.2015

    NP1 interacts and colocalizes with Kv7.2 at presynaptic terminals of excitatory synapses and axonal growth cones. A – D , Representative Western blots of immunoprecipitation eluates separated in 6% Tris-Glycine (for high molecular weight proteins) and 4–12% Bis-Tris gels (for low molecular weight proteins). TL, Total protein lysate; Syx, syntaxin. A , Both native Kv7.2 and syntaxin 1A, but not Kv7.3, coprecipitate with NP1 in total brain extracts. B , Both native Kv7.2 and NP1 coprecipitate with syntaxin in total brain extracts. C , D , Recombinant NP1 coprecipitates Kv7.2, but not syntaxin or Kv7.3, in 293T cells transfected with NP1, 5Myc-Kv7.2, 2HA-Kv7.3, and syntaxin 1A cDNAs. Kv7.2 and Kv7.3 were immunoprecipitated with antibodies against their respective Myc and HA tags. E , F , Immunofluorescence studies and confocal microscopy were performed in high-density ( E ) or low-density ( F ) isolated cortical neurons. E , Top, Confocal sections of 0.772 μm in the z -plane showing immunofluorescence of NP1, VGLUT1, Kv7.2, and negative control (omitting primary antibodies). Bottom, Colocalization (in white) of the excitatory presynaptic marker VGLUT1 (blue) with NP1 (green) and Kv7.2 (red) is shown in a single section with the corresponding orthogonal views of the stack of confocal sections. White arrows indicate sites of colocalization. F , NP1 (green) and KV7.2 (magenta) immunofluorescence and DIC images of an isolated cortical cultured neuron (1×) with its corresponding axonal growth cone highlighted in a white square box, shown in a confocal section of 0.772 μm in the z -plane at higher (5×) magnification. The negative control for primary antibodies is shown in another growth cone on the right. The image in the bottom is the merge of NP1 and Kv7.2 immunofluorescence images in the single confocal section obtained at 5× showing colocalization (white) of NP1 and Kv7.2 in the growth cone, with the corresponding orthogonal views from its respective stack of confocal sections. Images were acquired using restricted spectral emission wavelength ranges chosen to avoid crosstalk or bleed-through between the three different channels. Scale bar, 5 μm.
    Figure Legend Snippet: NP1 interacts and colocalizes with Kv7.2 at presynaptic terminals of excitatory synapses and axonal growth cones. A – D , Representative Western blots of immunoprecipitation eluates separated in 6% Tris-Glycine (for high molecular weight proteins) and 4–12% Bis-Tris gels (for low molecular weight proteins). TL, Total protein lysate; Syx, syntaxin. A , Both native Kv7.2 and syntaxin 1A, but not Kv7.3, coprecipitate with NP1 in total brain extracts. B , Both native Kv7.2 and NP1 coprecipitate with syntaxin in total brain extracts. C , D , Recombinant NP1 coprecipitates Kv7.2, but not syntaxin or Kv7.3, in 293T cells transfected with NP1, 5Myc-Kv7.2, 2HA-Kv7.3, and syntaxin 1A cDNAs. Kv7.2 and Kv7.3 were immunoprecipitated with antibodies against their respective Myc and HA tags. E , F , Immunofluorescence studies and confocal microscopy were performed in high-density ( E ) or low-density ( F ) isolated cortical neurons. E , Top, Confocal sections of 0.772 μm in the z -plane showing immunofluorescence of NP1, VGLUT1, Kv7.2, and negative control (omitting primary antibodies). Bottom, Colocalization (in white) of the excitatory presynaptic marker VGLUT1 (blue) with NP1 (green) and Kv7.2 (red) is shown in a single section with the corresponding orthogonal views of the stack of confocal sections. White arrows indicate sites of colocalization. F , NP1 (green) and KV7.2 (magenta) immunofluorescence and DIC images of an isolated cortical cultured neuron (1×) with its corresponding axonal growth cone highlighted in a white square box, shown in a confocal section of 0.772 μm in the z -plane at higher (5×) magnification. The negative control for primary antibodies is shown in another growth cone on the right. The image in the bottom is the merge of NP1 and Kv7.2 immunofluorescence images in the single confocal section obtained at 5× showing colocalization (white) of NP1 and Kv7.2 in the growth cone, with the corresponding orthogonal views from its respective stack of confocal sections. Images were acquired using restricted spectral emission wavelength ranges chosen to avoid crosstalk or bleed-through between the three different channels. Scale bar, 5 μm.

    Techniques Used: Western Blot, Immunoprecipitation, Molecular Weight, Recombinant, Transfection, Immunofluorescence, Confocal Microscopy, Isolation, Negative Control, Marker, Cell Culture

    Knockdown of NP1 reduces Kv7.2 surface expression and M-current. A , B , The proportion of Kv7.2 at the neuronal surface was determined by biotinylation (30 min at 4°C) of all surface proteins in cultured cortical neurons. The ratio of the surface over the total amount of receptor was determined by densitometric analysis and expressed as a percentage of control values. Only samples of biotinylated proteins lacking immunoreactivity to actin, which indicates contamination with intracellular proteins, were used for analyses. A , Representative Western blots of Kv7.2, Na-KATPase, and actin from total (T) and biotinylated (B) proteins. B , Quantitative analysis of the effect of gain and loss of NP1 on the surface/total levels of the Kv7.2 subunit of the voltage-gated Kv7/M K + channels. Values are mean ± SEM of shRdm ( n = 11), shNP1 ( n = 11), and pWPI-NP1 ( n = 5), from at least three independent experiments; * p
    Figure Legend Snippet: Knockdown of NP1 reduces Kv7.2 surface expression and M-current. A , B , The proportion of Kv7.2 at the neuronal surface was determined by biotinylation (30 min at 4°C) of all surface proteins in cultured cortical neurons. The ratio of the surface over the total amount of receptor was determined by densitometric analysis and expressed as a percentage of control values. Only samples of biotinylated proteins lacking immunoreactivity to actin, which indicates contamination with intracellular proteins, were used for analyses. A , Representative Western blots of Kv7.2, Na-KATPase, and actin from total (T) and biotinylated (B) proteins. B , Quantitative analysis of the effect of gain and loss of NP1 on the surface/total levels of the Kv7.2 subunit of the voltage-gated Kv7/M K + channels. Values are mean ± SEM of shRdm ( n = 11), shNP1 ( n = 11), and pWPI-NP1 ( n = 5), from at least three independent experiments; * p

    Techniques Used: Expressing, Cell Culture, Western Blot

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    Alomone Labs rabbit anti kv7 2
    NP1 interacts and colocalizes with <t>Kv7.2</t> at presynaptic terminals of excitatory synapses and axonal growth cones. A – D , Representative Western blots of immunoprecipitation eluates separated in 6% Tris-Glycine (for high molecular weight proteins) and 4–12% Bis-Tris gels (for low molecular weight proteins). TL, Total protein lysate; Syx, syntaxin. A , Both native Kv7.2 and syntaxin 1A, but not Kv7.3, coprecipitate with NP1 in total brain extracts. B , Both native Kv7.2 and NP1 coprecipitate with syntaxin in total brain extracts. C , D , Recombinant NP1 coprecipitates Kv7.2, but not syntaxin or Kv7.3, in 293T cells transfected with NP1, 5Myc-Kv7.2, 2HA-Kv7.3, and syntaxin 1A cDNAs. Kv7.2 and Kv7.3 were immunoprecipitated with antibodies against their respective Myc and HA tags. E , F , Immunofluorescence studies and confocal microscopy were performed in high-density ( E ) or low-density ( F ) isolated cortical neurons. E , Top, Confocal sections of 0.772 μm in the z -plane showing immunofluorescence of NP1, VGLUT1, Kv7.2, and negative control (omitting primary antibodies). Bottom, Colocalization (in white) of the excitatory presynaptic marker VGLUT1 (blue) with NP1 (green) and Kv7.2 (red) is shown in a single section with the corresponding orthogonal views of the stack of confocal sections. White arrows indicate sites of colocalization. F , NP1 (green) and KV7.2 (magenta) immunofluorescence and DIC images of an isolated cortical cultured neuron (1×) with its corresponding axonal growth cone highlighted in a white square box, shown in a confocal section of 0.772 μm in the z -plane at higher (5×) magnification. The negative control for primary antibodies is shown in another growth cone on the right. The image in the bottom is the merge of NP1 and Kv7.2 immunofluorescence images in the single confocal section obtained at 5× showing colocalization (white) of NP1 and Kv7.2 in the growth cone, with the corresponding orthogonal views from its respective stack of confocal sections. Images were acquired using restricted spectral emission wavelength ranges chosen to avoid crosstalk or bleed-through between the three different channels. Scale bar, 5 μm.
    Rabbit Anti Kv7 2, 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/rabbit anti kv7 2/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti kv7 2 - by Bioz Stars, 2022-08
    93/100 stars
      Buy from Supplier

    94
    Alomone Labs kv7 2
    Long-term drinking reduced SUMOylation levels of <t>Kv7.2</t> in the NAc. (a) Schematic depicting locations of the SUMOylation consensus sequences in rat Kv7.2 (orange circle and arrow indicate point of SUMO attachment). (b) Representative western blot images showing the expression of SUMOylated Kv7.2 in fractionated NAc tissue. (c) 72 hr withdrawal from alcohol decreased expression of SUMOylated Kv7.2 in the DSM but not the DRM fraction. (d) However, IAA exposure reduced the ratio of SUMOylated Kv7.2 to total Kv7.2 in the DRM fraction (* p
    Kv7 2, 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
    https://www.bioz.com/result/kv7 2/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    kv7 2 - by Bioz Stars, 2022-08
    94/100 stars
      Buy from Supplier

    93
    Alomone Labs rabbit anti kv7 2 antibody
    NP1 interacts and colocalizes with <t>Kv7.2</t> at presynaptic terminals of excitatory synapses and axonal growth cones. A – D , Representative Western blots of immunoprecipitation eluates separated in 6% Tris-Glycine (for high molecular weight proteins) and 4–12% Bis-Tris gels (for low molecular weight proteins). TL, Total protein lysate; Syx, syntaxin. A , Both native Kv7.2 and syntaxin 1A, but not Kv7.3, coprecipitate with NP1 in total brain extracts. B , Both native Kv7.2 and NP1 coprecipitate with syntaxin in total brain extracts. C , D , Recombinant NP1 coprecipitates Kv7.2, but not syntaxin or Kv7.3, in 293T cells transfected with NP1, 5Myc-Kv7.2, 2HA-Kv7.3, and syntaxin 1A cDNAs. Kv7.2 and Kv7.3 were immunoprecipitated with antibodies against their respective Myc and HA tags. E , F , Immunofluorescence studies and confocal microscopy were performed in high-density ( E ) or low-density ( F ) isolated cortical neurons. E , Top, Confocal sections of 0.772 μm in the z -plane showing immunofluorescence of NP1, VGLUT1, Kv7.2, and negative control (omitting primary antibodies). Bottom, Colocalization (in white) of the excitatory presynaptic marker VGLUT1 (blue) with NP1 (green) and Kv7.2 (red) is shown in a single section with the corresponding orthogonal views of the stack of confocal sections. White arrows indicate sites of colocalization. F , NP1 (green) and KV7.2 (magenta) immunofluorescence and DIC images of an isolated cortical cultured neuron (1×) with its corresponding axonal growth cone highlighted in a white square box, shown in a confocal section of 0.772 μm in the z -plane at higher (5×) magnification. The negative control for primary antibodies is shown in another growth cone on the right. The image in the bottom is the merge of NP1 and Kv7.2 immunofluorescence images in the single confocal section obtained at 5× showing colocalization (white) of NP1 and Kv7.2 in the growth cone, with the corresponding orthogonal views from its respective stack of confocal sections. Images were acquired using restricted spectral emission wavelength ranges chosen to avoid crosstalk or bleed-through between the three different channels. Scale bar, 5 μm.
    Rabbit Anti Kv7 2 Antibody, 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/rabbit anti kv7 2 antibody/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti kv7 2 antibody - by Bioz Stars, 2022-08
    93/100 stars
      Buy from Supplier

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    NP1 interacts and colocalizes with Kv7.2 at presynaptic terminals of excitatory synapses and axonal growth cones. A – D , Representative Western blots of immunoprecipitation eluates separated in 6% Tris-Glycine (for high molecular weight proteins) and 4–12% Bis-Tris gels (for low molecular weight proteins). TL, Total protein lysate; Syx, syntaxin. A , Both native Kv7.2 and syntaxin 1A, but not Kv7.3, coprecipitate with NP1 in total brain extracts. B , Both native Kv7.2 and NP1 coprecipitate with syntaxin in total brain extracts. C , D , Recombinant NP1 coprecipitates Kv7.2, but not syntaxin or Kv7.3, in 293T cells transfected with NP1, 5Myc-Kv7.2, 2HA-Kv7.3, and syntaxin 1A cDNAs. Kv7.2 and Kv7.3 were immunoprecipitated with antibodies against their respective Myc and HA tags. E , F , Immunofluorescence studies and confocal microscopy were performed in high-density ( E ) or low-density ( F ) isolated cortical neurons. E , Top, Confocal sections of 0.772 μm in the z -plane showing immunofluorescence of NP1, VGLUT1, Kv7.2, and negative control (omitting primary antibodies). Bottom, Colocalization (in white) of the excitatory presynaptic marker VGLUT1 (blue) with NP1 (green) and Kv7.2 (red) is shown in a single section with the corresponding orthogonal views of the stack of confocal sections. White arrows indicate sites of colocalization. F , NP1 (green) and KV7.2 (magenta) immunofluorescence and DIC images of an isolated cortical cultured neuron (1×) with its corresponding axonal growth cone highlighted in a white square box, shown in a confocal section of 0.772 μm in the z -plane at higher (5×) magnification. The negative control for primary antibodies is shown in another growth cone on the right. The image in the bottom is the merge of NP1 and Kv7.2 immunofluorescence images in the single confocal section obtained at 5× showing colocalization (white) of NP1 and Kv7.2 in the growth cone, with the corresponding orthogonal views from its respective stack of confocal sections. Images were acquired using restricted spectral emission wavelength ranges chosen to avoid crosstalk or bleed-through between the three different channels. Scale bar, 5 μm.

    Journal: The Journal of Neuroscience

    Article Title: Neuronal Pentraxin 1 Negatively Regulates Excitatory Synapse Density and Synaptic Plasticity

    doi: 10.1523/JNEUROSCI.2548-14.2015

    Figure Lengend Snippet: NP1 interacts and colocalizes with Kv7.2 at presynaptic terminals of excitatory synapses and axonal growth cones. A – D , Representative Western blots of immunoprecipitation eluates separated in 6% Tris-Glycine (for high molecular weight proteins) and 4–12% Bis-Tris gels (for low molecular weight proteins). TL, Total protein lysate; Syx, syntaxin. A , Both native Kv7.2 and syntaxin 1A, but not Kv7.3, coprecipitate with NP1 in total brain extracts. B , Both native Kv7.2 and NP1 coprecipitate with syntaxin in total brain extracts. C , D , Recombinant NP1 coprecipitates Kv7.2, but not syntaxin or Kv7.3, in 293T cells transfected with NP1, 5Myc-Kv7.2, 2HA-Kv7.3, and syntaxin 1A cDNAs. Kv7.2 and Kv7.3 were immunoprecipitated with antibodies against their respective Myc and HA tags. E , F , Immunofluorescence studies and confocal microscopy were performed in high-density ( E ) or low-density ( F ) isolated cortical neurons. E , Top, Confocal sections of 0.772 μm in the z -plane showing immunofluorescence of NP1, VGLUT1, Kv7.2, and negative control (omitting primary antibodies). Bottom, Colocalization (in white) of the excitatory presynaptic marker VGLUT1 (blue) with NP1 (green) and Kv7.2 (red) is shown in a single section with the corresponding orthogonal views of the stack of confocal sections. White arrows indicate sites of colocalization. F , NP1 (green) and KV7.2 (magenta) immunofluorescence and DIC images of an isolated cortical cultured neuron (1×) with its corresponding axonal growth cone highlighted in a white square box, shown in a confocal section of 0.772 μm in the z -plane at higher (5×) magnification. The negative control for primary antibodies is shown in another growth cone on the right. The image in the bottom is the merge of NP1 and Kv7.2 immunofluorescence images in the single confocal section obtained at 5× showing colocalization (white) of NP1 and Kv7.2 in the growth cone, with the corresponding orthogonal views from its respective stack of confocal sections. Images were acquired using restricted spectral emission wavelength ranges chosen to avoid crosstalk or bleed-through between the three different channels. Scale bar, 5 μm.

    Article Snippet: We used the following antibodies: mouse anti-NP1 antibody (1:1000; Becton Dickinson), mouse anti-synaptophysin, clone SY38 (1:1000), mouse anti-PSD95 (1:2000; both from Millipore Bioscience Research Reagents), rabbit anti-Kv7.2 (1:200; Alomone Labs), mouse anti-syntaxin 1A (HPC-1 clone, 1:2000; Sigma-Aldrich), mouse anti-HA (1:3000, Covance), and mouse anti-Myc (9E10 clone, 1:5000; Sigma-Aldrich).

    Techniques: Western Blot, Immunoprecipitation, Molecular Weight, Recombinant, Transfection, Immunofluorescence, Confocal Microscopy, Isolation, Negative Control, Marker, Cell Culture

    Knockdown of NP1 reduces Kv7.2 surface expression and M-current. A , B , The proportion of Kv7.2 at the neuronal surface was determined by biotinylation (30 min at 4°C) of all surface proteins in cultured cortical neurons. The ratio of the surface over the total amount of receptor was determined by densitometric analysis and expressed as a percentage of control values. Only samples of biotinylated proteins lacking immunoreactivity to actin, which indicates contamination with intracellular proteins, were used for analyses. A , Representative Western blots of Kv7.2, Na-KATPase, and actin from total (T) and biotinylated (B) proteins. B , Quantitative analysis of the effect of gain and loss of NP1 on the surface/total levels of the Kv7.2 subunit of the voltage-gated Kv7/M K + channels. Values are mean ± SEM of shRdm ( n = 11), shNP1 ( n = 11), and pWPI-NP1 ( n = 5), from at least three independent experiments; * p

    Journal: The Journal of Neuroscience

    Article Title: Neuronal Pentraxin 1 Negatively Regulates Excitatory Synapse Density and Synaptic Plasticity

    doi: 10.1523/JNEUROSCI.2548-14.2015

    Figure Lengend Snippet: Knockdown of NP1 reduces Kv7.2 surface expression and M-current. A , B , The proportion of Kv7.2 at the neuronal surface was determined by biotinylation (30 min at 4°C) of all surface proteins in cultured cortical neurons. The ratio of the surface over the total amount of receptor was determined by densitometric analysis and expressed as a percentage of control values. Only samples of biotinylated proteins lacking immunoreactivity to actin, which indicates contamination with intracellular proteins, were used for analyses. A , Representative Western blots of Kv7.2, Na-KATPase, and actin from total (T) and biotinylated (B) proteins. B , Quantitative analysis of the effect of gain and loss of NP1 on the surface/total levels of the Kv7.2 subunit of the voltage-gated Kv7/M K + channels. Values are mean ± SEM of shRdm ( n = 11), shNP1 ( n = 11), and pWPI-NP1 ( n = 5), from at least three independent experiments; * p

    Article Snippet: We used the following antibodies: mouse anti-NP1 antibody (1:1000; Becton Dickinson), mouse anti-synaptophysin, clone SY38 (1:1000), mouse anti-PSD95 (1:2000; both from Millipore Bioscience Research Reagents), rabbit anti-Kv7.2 (1:200; Alomone Labs), mouse anti-syntaxin 1A (HPC-1 clone, 1:2000; Sigma-Aldrich), mouse anti-HA (1:3000, Covance), and mouse anti-Myc (9E10 clone, 1:5000; Sigma-Aldrich).

    Techniques: Expressing, Cell Culture, Western Blot

    Long-term drinking reduced SUMOylation levels of Kv7.2 in the NAc. (a) Schematic depicting locations of the SUMOylation consensus sequences in rat Kv7.2 (orange circle and arrow indicate point of SUMO attachment). (b) Representative western blot images showing the expression of SUMOylated Kv7.2 in fractionated NAc tissue. (c) 72 hr withdrawal from alcohol decreased expression of SUMOylated Kv7.2 in the DSM but not the DRM fraction. (d) However, IAA exposure reduced the ratio of SUMOylated Kv7.2 to total Kv7.2 in the DRM fraction (* p

    Journal: Addiction biology

    Article Title: Kv7 Channels in the Nucleus Accumbens are altered by Chronic Drinking and are Targets for Reducing Alcohol Consumption

    doi: 10.1111/adb.12279

    Figure Lengend Snippet: Long-term drinking reduced SUMOylation levels of Kv7.2 in the NAc. (a) Schematic depicting locations of the SUMOylation consensus sequences in rat Kv7.2 (orange circle and arrow indicate point of SUMO attachment). (b) Representative western blot images showing the expression of SUMOylated Kv7.2 in fractionated NAc tissue. (c) 72 hr withdrawal from alcohol decreased expression of SUMOylated Kv7.2 in the DSM but not the DRM fraction. (d) However, IAA exposure reduced the ratio of SUMOylated Kv7.2 to total Kv7.2 in the DRM fraction (* p

    Article Snippet: The antibodies used in this study were Kv7.2 (#APC-050; Alomone Labs; Jerusalem, Israel), AKAP150 (#07-210; EMD Millipore; Billerica, MA), and SGK1 (#S5188; Sigma-Aldrich Co).

    Techniques: Western Blot, Expressing

    Long-term drinking altered Kv7.2 surface trafficking in the NAc. (a) Representative western blot images showing the relative expression patterns of Kv7.2, AKAP150, and SGK1 in the DRM, IC, and DSM fractions in tissue targeting the NAc core. (b) Representative western blot images of Kv7.2, AKAP150, and SGK1 expression in the NAc of alcohol naïve and IAA rats after 72 hr withdrawal from alcohol. (c) Kv7.2 expression is increased in the DRM of IAA rats and decreased in the DSM. There is no effect of alcohol exposure on the expression of (d) AKAP150 in the DRM or (e) SGK1 in the IC fraction (* p

    Journal: Addiction biology

    Article Title: Kv7 Channels in the Nucleus Accumbens are altered by Chronic Drinking and are Targets for Reducing Alcohol Consumption

    doi: 10.1111/adb.12279

    Figure Lengend Snippet: Long-term drinking altered Kv7.2 surface trafficking in the NAc. (a) Representative western blot images showing the relative expression patterns of Kv7.2, AKAP150, and SGK1 in the DRM, IC, and DSM fractions in tissue targeting the NAc core. (b) Representative western blot images of Kv7.2, AKAP150, and SGK1 expression in the NAc of alcohol naïve and IAA rats after 72 hr withdrawal from alcohol. (c) Kv7.2 expression is increased in the DRM of IAA rats and decreased in the DSM. There is no effect of alcohol exposure on the expression of (d) AKAP150 in the DRM or (e) SGK1 in the IC fraction (* p

    Article Snippet: The antibodies used in this study were Kv7.2 (#APC-050; Alomone Labs; Jerusalem, Israel), AKAP150 (#07-210; EMD Millipore; Billerica, MA), and SGK1 (#S5188; Sigma-Aldrich Co).

    Techniques: Western Blot, Expressing

    Inhibition of G9a activity normalizes K + channel gene expression in the DRG diminished by nerve injury ( a-d) Effects of intrathecal treatments with vehicle (n = 10), SAHA (50 μg, n = 9), UNC0638 (10 μg, n = 8), GSK503 (5 μg, n = 10), SAHA plus GSK503 (n = 8), SAHA plus UNC0638 (n = 9) or UNC0638 plus GSK503 (n = 8) on the mRNA levels of Kcna4 ( a ), Kcnd2 ( b ), Kcnq2 ( c ), and Kcnma1 ( d ) in the DRG obtained from SNL rats 28 days after surgery. Data from sham control rats were plotted as the control (n = 6 rats). ( e,f ) Effects of nerve injury and UNC0638 on the protein levels of Kv1.4, Kv4.2, Kv7.2 and BKα1 in the L5 and L6 DRG (n = 6 rats in each group). ( g,h ) Effect of G9a-specific siRNA on the G9a and H3K9me2 protein levels in the DRG obtained from SNL rats 24 h after the last injection (n = 5 in each group). ( i,j ) Effects of G9a-specific siRNA on the mRNA levels of G9a, Ezh2, Kcna4, Kcnd2, Kcnq2 and Kcnma1 in the DRG obtained from SNL ( i ) and sham control ( j ) rats 24 h after the last injection (n = 10 in each group). Data are presented as mean ± s.e.m. Statistical analysis was performed using two-way ANOVA followed by Bonferroni’s post hoc tests ( a-d ), one-way ANOVA ( f,h,i ), or Mann-Whitney test ( j ). * P

    Journal: Nature neuroscience

    Article Title: G9a Is Essential for Epigenetic Silencing of K+ Channel Genes in Acute-to-Chronic Pain Transition

    doi: 10.1038/nn.4165

    Figure Lengend Snippet: Inhibition of G9a activity normalizes K + channel gene expression in the DRG diminished by nerve injury ( a-d) Effects of intrathecal treatments with vehicle (n = 10), SAHA (50 μg, n = 9), UNC0638 (10 μg, n = 8), GSK503 (5 μg, n = 10), SAHA plus GSK503 (n = 8), SAHA plus UNC0638 (n = 9) or UNC0638 plus GSK503 (n = 8) on the mRNA levels of Kcna4 ( a ), Kcnd2 ( b ), Kcnq2 ( c ), and Kcnma1 ( d ) in the DRG obtained from SNL rats 28 days after surgery. Data from sham control rats were plotted as the control (n = 6 rats). ( e,f ) Effects of nerve injury and UNC0638 on the protein levels of Kv1.4, Kv4.2, Kv7.2 and BKα1 in the L5 and L6 DRG (n = 6 rats in each group). ( g,h ) Effect of G9a-specific siRNA on the G9a and H3K9me2 protein levels in the DRG obtained from SNL rats 24 h after the last injection (n = 5 in each group). ( i,j ) Effects of G9a-specific siRNA on the mRNA levels of G9a, Ezh2, Kcna4, Kcnd2, Kcnq2 and Kcnma1 in the DRG obtained from SNL ( i ) and sham control ( j ) rats 24 h after the last injection (n = 10 in each group). Data are presented as mean ± s.e.m. Statistical analysis was performed using two-way ANOVA followed by Bonferroni’s post hoc tests ( a-d ), one-way ANOVA ( f,h,i ), or Mann-Whitney test ( j ). * P

    Article Snippet: The membranes were treated with 5% bovine serum albumin in Tris buffer containing Tween 20 (TNT) for 2 h and then incubated with one of the following primary antibodies overnight at 4°C: Kv1.4 (catalog #05-409, Upstate Biotechnology), Kv4.2 (catalog #APC-023, Alomone Labs), Kv7.2 (catalog #APC-050, Alomone Labs), BKα1 (catalog #APC-021, Alomone Labs), acetyl-H3 (catalog #06-599, Millipore), H3K9ac (catalog #39917, Active Motif), H3K27me3 (catalog #9733, Cell Signaling Technology), histone H3 (catalog #9715, Cell Signaling Technology), H3K9me2 (catalog #ab1220, Abcam), G9a (catalog #09-071, Millipore), HDAC1 (catalog #IMG-337, Imgenex), and HDAC2, HDAC4, HDAC5 and EZH2 (catalog #2540, #2072, #2082 and #5246, respectively; Cell Signaling Technology).

    Techniques: Inhibition, Activity Assay, Expressing, Injection, MANN-WHITNEY

    NP1 interacts and colocalizes with Kv7.2 at presynaptic terminals of excitatory synapses and axonal growth cones. A – D , Representative Western blots of immunoprecipitation eluates separated in 6% Tris-Glycine (for high molecular weight proteins) and 4–12% Bis-Tris gels (for low molecular weight proteins). TL, Total protein lysate; Syx, syntaxin. A , Both native Kv7.2 and syntaxin 1A, but not Kv7.3, coprecipitate with NP1 in total brain extracts. B , Both native Kv7.2 and NP1 coprecipitate with syntaxin in total brain extracts. C , D , Recombinant NP1 coprecipitates Kv7.2, but not syntaxin or Kv7.3, in 293T cells transfected with NP1, 5Myc-Kv7.2, 2HA-Kv7.3, and syntaxin 1A cDNAs. Kv7.2 and Kv7.3 were immunoprecipitated with antibodies against their respective Myc and HA tags. E , F , Immunofluorescence studies and confocal microscopy were performed in high-density ( E ) or low-density ( F ) isolated cortical neurons. E , Top, Confocal sections of 0.772 μm in the z -plane showing immunofluorescence of NP1, VGLUT1, Kv7.2, and negative control (omitting primary antibodies). Bottom, Colocalization (in white) of the excitatory presynaptic marker VGLUT1 (blue) with NP1 (green) and Kv7.2 (red) is shown in a single section with the corresponding orthogonal views of the stack of confocal sections. White arrows indicate sites of colocalization. F , NP1 (green) and KV7.2 (magenta) immunofluorescence and DIC images of an isolated cortical cultured neuron (1×) with its corresponding axonal growth cone highlighted in a white square box, shown in a confocal section of 0.772 μm in the z -plane at higher (5×) magnification. The negative control for primary antibodies is shown in another growth cone on the right. The image in the bottom is the merge of NP1 and Kv7.2 immunofluorescence images in the single confocal section obtained at 5× showing colocalization (white) of NP1 and Kv7.2 in the growth cone, with the corresponding orthogonal views from its respective stack of confocal sections. Images were acquired using restricted spectral emission wavelength ranges chosen to avoid crosstalk or bleed-through between the three different channels. Scale bar, 5 μm.

    Journal: The Journal of Neuroscience

    Article Title: Neuronal Pentraxin 1 Negatively Regulates Excitatory Synapse Density and Synaptic Plasticity

    doi: 10.1523/JNEUROSCI.2548-14.2015

    Figure Lengend Snippet: NP1 interacts and colocalizes with Kv7.2 at presynaptic terminals of excitatory synapses and axonal growth cones. A – D , Representative Western blots of immunoprecipitation eluates separated in 6% Tris-Glycine (for high molecular weight proteins) and 4–12% Bis-Tris gels (for low molecular weight proteins). TL, Total protein lysate; Syx, syntaxin. A , Both native Kv7.2 and syntaxin 1A, but not Kv7.3, coprecipitate with NP1 in total brain extracts. B , Both native Kv7.2 and NP1 coprecipitate with syntaxin in total brain extracts. C , D , Recombinant NP1 coprecipitates Kv7.2, but not syntaxin or Kv7.3, in 293T cells transfected with NP1, 5Myc-Kv7.2, 2HA-Kv7.3, and syntaxin 1A cDNAs. Kv7.2 and Kv7.3 were immunoprecipitated with antibodies against their respective Myc and HA tags. E , F , Immunofluorescence studies and confocal microscopy were performed in high-density ( E ) or low-density ( F ) isolated cortical neurons. E , Top, Confocal sections of 0.772 μm in the z -plane showing immunofluorescence of NP1, VGLUT1, Kv7.2, and negative control (omitting primary antibodies). Bottom, Colocalization (in white) of the excitatory presynaptic marker VGLUT1 (blue) with NP1 (green) and Kv7.2 (red) is shown in a single section with the corresponding orthogonal views of the stack of confocal sections. White arrows indicate sites of colocalization. F , NP1 (green) and KV7.2 (magenta) immunofluorescence and DIC images of an isolated cortical cultured neuron (1×) with its corresponding axonal growth cone highlighted in a white square box, shown in a confocal section of 0.772 μm in the z -plane at higher (5×) magnification. The negative control for primary antibodies is shown in another growth cone on the right. The image in the bottom is the merge of NP1 and Kv7.2 immunofluorescence images in the single confocal section obtained at 5× showing colocalization (white) of NP1 and Kv7.2 in the growth cone, with the corresponding orthogonal views from its respective stack of confocal sections. Images were acquired using restricted spectral emission wavelength ranges chosen to avoid crosstalk or bleed-through between the three different channels. Scale bar, 5 μm.

    Article Snippet: The bound biotinylated proteins were eluted by incubating for 1 h at room temperature with SDS-PAGE buffer (62.5 m m Tris-HCl, pH 6.8, 1% SDS, 10% glycerol, and 50 m m DTT) followed by centrifugation for 2 min at 1000 × g. Biotinylated proteins were separated in 8% SDS-PAGE and transferred to PVDF membranes (Millipore), which were probed with one of the following antibodies: rabbit anti-Kv7.2 antibody (1:2000; Alomone Labs), rabbit anti-GluA1 antibody (1:1000; Millipore), mouse monoclonal anti-GluA2 antibody (1:1000; Neuromab), and mouse monoclonal anti-GluN1 (1:2000; Becton Dickinson) as described above.

    Techniques: Western Blot, Immunoprecipitation, Molecular Weight, Recombinant, Transfection, Immunofluorescence, Confocal Microscopy, Isolation, Negative Control, Marker, Cell Culture

    Knockdown of NP1 reduces Kv7.2 surface expression and M-current. A , B , The proportion of Kv7.2 at the neuronal surface was determined by biotinylation (30 min at 4°C) of all surface proteins in cultured cortical neurons. The ratio of the surface over the total amount of receptor was determined by densitometric analysis and expressed as a percentage of control values. Only samples of biotinylated proteins lacking immunoreactivity to actin, which indicates contamination with intracellular proteins, were used for analyses. A , Representative Western blots of Kv7.2, Na-KATPase, and actin from total (T) and biotinylated (B) proteins. B , Quantitative analysis of the effect of gain and loss of NP1 on the surface/total levels of the Kv7.2 subunit of the voltage-gated Kv7/M K + channels. Values are mean ± SEM of shRdm ( n = 11), shNP1 ( n = 11), and pWPI-NP1 ( n = 5), from at least three independent experiments; * p

    Journal: The Journal of Neuroscience

    Article Title: Neuronal Pentraxin 1 Negatively Regulates Excitatory Synapse Density and Synaptic Plasticity

    doi: 10.1523/JNEUROSCI.2548-14.2015

    Figure Lengend Snippet: Knockdown of NP1 reduces Kv7.2 surface expression and M-current. A , B , The proportion of Kv7.2 at the neuronal surface was determined by biotinylation (30 min at 4°C) of all surface proteins in cultured cortical neurons. The ratio of the surface over the total amount of receptor was determined by densitometric analysis and expressed as a percentage of control values. Only samples of biotinylated proteins lacking immunoreactivity to actin, which indicates contamination with intracellular proteins, were used for analyses. A , Representative Western blots of Kv7.2, Na-KATPase, and actin from total (T) and biotinylated (B) proteins. B , Quantitative analysis of the effect of gain and loss of NP1 on the surface/total levels of the Kv7.2 subunit of the voltage-gated Kv7/M K + channels. Values are mean ± SEM of shRdm ( n = 11), shNP1 ( n = 11), and pWPI-NP1 ( n = 5), from at least three independent experiments; * p

    Article Snippet: The bound biotinylated proteins were eluted by incubating for 1 h at room temperature with SDS-PAGE buffer (62.5 m m Tris-HCl, pH 6.8, 1% SDS, 10% glycerol, and 50 m m DTT) followed by centrifugation for 2 min at 1000 × g. Biotinylated proteins were separated in 8% SDS-PAGE and transferred to PVDF membranes (Millipore), which were probed with one of the following antibodies: rabbit anti-Kv7.2 antibody (1:2000; Alomone Labs), rabbit anti-GluA1 antibody (1:1000; Millipore), mouse monoclonal anti-GluA2 antibody (1:1000; Neuromab), and mouse monoclonal anti-GluN1 (1:2000; Becton Dickinson) as described above.

    Techniques: Expressing, Cell Culture, Western Blot