kcnk10  (Alomone Labs)


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    Structured Review

    Alomone Labs kcnk10
    Kcnk10, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 86 stars, based on 1 article reviews
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    kcnk10 - by Bioz Stars, 2021-12
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    Alomone Labs rabbit anti trek2 antibody
    A , <t>TREK2</t> antibody characterization. Top, GFP fluorescence or interference contrast (IC) shows HEK cells, transfected with plasmids for six rat K2P channels: TREK2, TWIK1, TREK1, TASK3, THIK2, and THIK1 (all subcloned in pCMV-SPORT vector). Bottom, Same fields show immunostaining for TREK2 that was intrinsically very low (GFP alone), increased substantially after transfection with the chimeric TREK2-GFP and with TREK2, but remained very low after transfection with all non-TREK2 K2P channels tested. This antibody therefore selectively recognizes TREK2 and not the other K2P channel tested, including TREK1, structurally the closest related to TREK2. Scale bar, 40 μm. B–D , TREK2 in DRG neurons is selectively expressed in IB4 + neurons. B , Triple fluorescence immunocytochemistry shows TREK2 (blue) to be highly colocalized with IB4 binding (red). RT97 (green) is a marker for neurofilament (against a highly phosphorylated epitope on NF200) that selectively stains DRG somata with A-fibers (i.e., myelinated). TREK2 was only visible in neurofilament-poor (NF-poor, thus C-fiber) neurons, as shown by lack of (green) NF staining in TREK2 + neurons. TREK2 staining is thus in IB4 + DRG neurons with C-fibers. Scale bar, 20 μm. C , D , %intensities refers to cytoplasmic immunointensities expressed as a percentage of maximum staining above background for that marker (see Materials and Methods); gray band(s) show(s) negative staining for TREK2 (
    Rabbit Anti Trek2 Antibody, 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
    https://www.bioz.com/result/rabbit anti trek2 antibody/product/Alomone Labs
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti trek2 antibody - by Bioz Stars, 2021-12
    92/100 stars
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    86
    Alomone Labs polyclonal rabbit anti trek2 antibody
    A , <t>TREK2</t> antibody characterization. Top, GFP fluorescence or interference contrast (IC) shows HEK cells, transfected with plasmids for six rat K2P channels: TREK2, TWIK1, TREK1, TASK3, THIK2, and THIK1 (all subcloned in pCMV-SPORT vector). Bottom, Same fields show immunostaining for TREK2 that was intrinsically very low (GFP alone), increased substantially after transfection with the chimeric TREK2-GFP and with TREK2, but remained very low after transfection with all non-TREK2 K2P channels tested. This antibody therefore selectively recognizes TREK2 and not the other K2P channel tested, including TREK1, structurally the closest related to TREK2. Scale bar, 40 μm. B–D , TREK2 in DRG neurons is selectively expressed in IB4 + neurons. B , Triple fluorescence immunocytochemistry shows TREK2 (blue) to be highly colocalized with IB4 binding (red). RT97 (green) is a marker for neurofilament (against a highly phosphorylated epitope on NF200) that selectively stains DRG somata with A-fibers (i.e., myelinated). TREK2 was only visible in neurofilament-poor (NF-poor, thus C-fiber) neurons, as shown by lack of (green) NF staining in TREK2 + neurons. TREK2 staining is thus in IB4 + DRG neurons with C-fibers. Scale bar, 20 μm. C , D , %intensities refers to cytoplasmic immunointensities expressed as a percentage of maximum staining above background for that marker (see Materials and Methods); gray band(s) show(s) negative staining for TREK2 (
    Polyclonal Rabbit Anti Trek2 Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/polyclonal rabbit anti trek2 antibody/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    polyclonal rabbit anti trek2 antibody - by Bioz Stars, 2021-12
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    A , TREK2 antibody characterization. Top, GFP fluorescence or interference contrast (IC) shows HEK cells, transfected with plasmids for six rat K2P channels: TREK2, TWIK1, TREK1, TASK3, THIK2, and THIK1 (all subcloned in pCMV-SPORT vector). Bottom, Same fields show immunostaining for TREK2 that was intrinsically very low (GFP alone), increased substantially after transfection with the chimeric TREK2-GFP and with TREK2, but remained very low after transfection with all non-TREK2 K2P channels tested. This antibody therefore selectively recognizes TREK2 and not the other K2P channel tested, including TREK1, structurally the closest related to TREK2. Scale bar, 40 μm. B–D , TREK2 in DRG neurons is selectively expressed in IB4 + neurons. B , Triple fluorescence immunocytochemistry shows TREK2 (blue) to be highly colocalized with IB4 binding (red). RT97 (green) is a marker for neurofilament (against a highly phosphorylated epitope on NF200) that selectively stains DRG somata with A-fibers (i.e., myelinated). TREK2 was only visible in neurofilament-poor (NF-poor, thus C-fiber) neurons, as shown by lack of (green) NF staining in TREK2 + neurons. TREK2 staining is thus in IB4 + DRG neurons with C-fibers. Scale bar, 20 μm. C , D , %intensities refers to cytoplasmic immunointensities expressed as a percentage of maximum staining above background for that marker (see Materials and Methods); gray band(s) show(s) negative staining for TREK2 (

    Journal: The Journal of Neuroscience

    Article Title: TREK2 Expressed Selectively in IB4-Binding C-Fiber Nociceptors Hyperpolarizes Their Membrane Potentials and Limits Spontaneous Pain

    doi: 10.1523/JNEUROSCI.4528-13.2014

    Figure Lengend Snippet: A , TREK2 antibody characterization. Top, GFP fluorescence or interference contrast (IC) shows HEK cells, transfected with plasmids for six rat K2P channels: TREK2, TWIK1, TREK1, TASK3, THIK2, and THIK1 (all subcloned in pCMV-SPORT vector). Bottom, Same fields show immunostaining for TREK2 that was intrinsically very low (GFP alone), increased substantially after transfection with the chimeric TREK2-GFP and with TREK2, but remained very low after transfection with all non-TREK2 K2P channels tested. This antibody therefore selectively recognizes TREK2 and not the other K2P channel tested, including TREK1, structurally the closest related to TREK2. Scale bar, 40 μm. B–D , TREK2 in DRG neurons is selectively expressed in IB4 + neurons. B , Triple fluorescence immunocytochemistry shows TREK2 (blue) to be highly colocalized with IB4 binding (red). RT97 (green) is a marker for neurofilament (against a highly phosphorylated epitope on NF200) that selectively stains DRG somata with A-fibers (i.e., myelinated). TREK2 was only visible in neurofilament-poor (NF-poor, thus C-fiber) neurons, as shown by lack of (green) NF staining in TREK2 + neurons. TREK2 staining is thus in IB4 + DRG neurons with C-fibers. Scale bar, 20 μm. C , D , %intensities refers to cytoplasmic immunointensities expressed as a percentage of maximum staining above background for that marker (see Materials and Methods); gray band(s) show(s) negative staining for TREK2 (

    Article Snippet: Three days after transfection, the HEK cell cultures expressing the different K2P channels were washed with PBS, fixed with 4% paraformaldehyde in 4.2% sucrose (in PBS, pH 7.2) for 10 min at room temperature, rinsed with PBS, and processed for TREK2 immunocytochemistry with the rabbit anti-TREK2 antibody from Alomone Labs as described above.

    Techniques: Fluorescence, Transfection, Plasmid Preparation, Immunostaining, Immunocytochemistry, Binding Assay, Marker, Staining, Negative Staining

    Axotomy 7 d earlier decreased Nav1.8 and Nav1.7 as well as TREK2 expression. A , Blinded subjective scoring (0 = no visible staining; 7 = maximum staining intensity) of whole cytoplasmic Nav1.8 immunointensities in L5 DRG sections from 4 normal (N) and 4 axotomized rats ipsilaterally (Ax). Medians plus interquartile ranges are shown. The median decreased significantly after axotomy (Mann–Whitney test). ** p

    Journal: The Journal of Neuroscience

    Article Title: TREK2 Expressed Selectively in IB4-Binding C-Fiber Nociceptors Hyperpolarizes Their Membrane Potentials and Limits Spontaneous Pain

    doi: 10.1523/JNEUROSCI.4528-13.2014

    Figure Lengend Snippet: Axotomy 7 d earlier decreased Nav1.8 and Nav1.7 as well as TREK2 expression. A , Blinded subjective scoring (0 = no visible staining; 7 = maximum staining intensity) of whole cytoplasmic Nav1.8 immunointensities in L5 DRG sections from 4 normal (N) and 4 axotomized rats ipsilaterally (Ax). Medians plus interquartile ranges are shown. The median decreased significantly after axotomy (Mann–Whitney test). ** p

    Article Snippet: Three days after transfection, the HEK cell cultures expressing the different K2P channels were washed with PBS, fixed with 4% paraformaldehyde in 4.2% sucrose (in PBS, pH 7.2) for 10 min at room temperature, rinsed with PBS, and processed for TREK2 immunocytochemistry with the rabbit anti-TREK2 antibody from Alomone Labs as described above.

    Techniques: Expressing, Staining, MANN-WHITNEY

    In vivo , TREK2 is in small C-fiber nociceptors; their Ems relate to TREK2 levels. A–G , DRG neurons injected in vivo with fluorescent dye after Em recording and sensory receptor property identification (see Materials and Methods). A , Examples (arrows) of fluorescent dye-labeled neurons before (top) and after (bottom) TREK2 ABC immunocytochemistry. TREK2%intensities (see Materials and Methods) were determined at edge-, mid-, and inner- (perinuclear) cytoplasm ( B , inset). The two C-fiber nociceptors (C-Nocis) have (left) a −52 mV Em and strong cytoplasmic, including edge, TREK2 intensity; then (mid-left) a −42 mV Em, with weak edge- but stronger mid- and inner-TREK2. The Aδ nociceptor (mid-right) and Aα/β LTM (far right) were TREK2 − (see Materials and Methods and below). Scale bar, 20 μm. B–G , Cytoplasmic edge- and mid-TREK2%intensities of ≥25% (of maximum cytoplasmic staining; see Materials and Methods) were deemed positive. The negative staining (

    Journal: The Journal of Neuroscience

    Article Title: TREK2 Expressed Selectively in IB4-Binding C-Fiber Nociceptors Hyperpolarizes Their Membrane Potentials and Limits Spontaneous Pain

    doi: 10.1523/JNEUROSCI.4528-13.2014

    Figure Lengend Snippet: In vivo , TREK2 is in small C-fiber nociceptors; their Ems relate to TREK2 levels. A–G , DRG neurons injected in vivo with fluorescent dye after Em recording and sensory receptor property identification (see Materials and Methods). A , Examples (arrows) of fluorescent dye-labeled neurons before (top) and after (bottom) TREK2 ABC immunocytochemistry. TREK2%intensities (see Materials and Methods) were determined at edge-, mid-, and inner- (perinuclear) cytoplasm ( B , inset). The two C-fiber nociceptors (C-Nocis) have (left) a −52 mV Em and strong cytoplasmic, including edge, TREK2 intensity; then (mid-left) a −42 mV Em, with weak edge- but stronger mid- and inner-TREK2. The Aδ nociceptor (mid-right) and Aα/β LTM (far right) were TREK2 − (see Materials and Methods and below). Scale bar, 20 μm. B–G , Cytoplasmic edge- and mid-TREK2%intensities of ≥25% (of maximum cytoplasmic staining; see Materials and Methods) were deemed positive. The negative staining (

    Article Snippet: Three days after transfection, the HEK cell cultures expressing the different K2P channels were washed with PBS, fixed with 4% paraformaldehyde in 4.2% sucrose (in PBS, pH 7.2) for 10 min at room temperature, rinsed with PBS, and processed for TREK2 immunocytochemistry with the rabbit anti-TREK2 antibody from Alomone Labs as described above.

    Techniques: In Vivo, Injection, Labeling, Immunocytochemistry, Staining, Negative Staining

    TREK2 knockdown in vivo decreased SFL 1 d after CFA. Ipsilateral SFL in rats 1 d after an intradermal plantar CFA injection in one hind foot in the same region as an intradermal plantar injection of either TREK2 siRNA or scrambled siRNA given 3 d earlier. Numbers in B and C (superscripts) and D and E (beside data points) relate to rats from which tissue in the lanes of A was obtained; these numbers are included to enable comparison of TREK2 protein expression with other data from the same rats. A , TREK2 Western blot of skin tissue, at the site of scr (1, 2) or siRNA (3, 4) injection 4 d previously, and 1 d after either saline (1, 3) or CFA injection (2, 4) at the same site. Top, The lower TREK2 band (MW ∼50 kDa, the band that we and others find in DRG tissues) was almost eliminated by siRNA, not scr, treatment, regardless of whether rats were injected with CFA or saline. The upper band was not reduced after siRNA. The MWs of these bands are consistent with them being two of the three known rat TREK2 isoforms (see Results and Discussion). Bottom, α-Tubulin loading controls. B , TREK2 knockdown by TREK2 siRNA in nerve fibers and epidermis (Ep) in plantar skin. Left, after scr siRNA injection. Right, after TREK2 siRNA injection. Both treatments were followed by CFA injection. Tissue was taken 4 d after scr/siRNA treatment (1 d after CFA). Triple fluorescence staining for β-tubulin III (blue, to stain nerve fibers), IB4 binding (green), and TREK2 (red). White dashed lines indicate junctions between epidermis (above) and dermis (below). In the merged images, colocalization of all three markers shows as white or very pale (bottom left image). After siRNA, red TREK2 staining in epidermis and in groups of nerve fibers (shown as blue and/or green) was reduced, showing substantial knockdown in vivo of TREK2 near the siRNA injection site. Scale bar, 25 μm. C , TREK2 knockdown by local TREK2 siRNA in the nerve innervating plantar skin. This knockdown, seen 5–10 mm from the siRNA injection site, occurred whether rats were subsequently treated with saline or CFA. There was decreased fiber TREK2 immunostaining (red) in TREK2 siRNA, not scr siRNA, treated rats, so that IB4 + fibers (green) showed little/no TREK2 (red). Scale bar, 50 μm. D , E , SFL after CFA increased after TREK2 siRNA compared with scr control. Ipsilateral SFLd ( D ) or SFLn ( E ) after siRNA C compared with scr treatment did not increase before saline injection (day 3, columns 1 and 2) or after saline injection (day 4, columns 3 and 4), or before CFA injection (day 3, columns 5 and 6). However, after CFA, it caused increased SFLd ( D ) and SFLn ( E ) in both scr ( p

    Journal: The Journal of Neuroscience

    Article Title: TREK2 Expressed Selectively in IB4-Binding C-Fiber Nociceptors Hyperpolarizes Their Membrane Potentials and Limits Spontaneous Pain

    doi: 10.1523/JNEUROSCI.4528-13.2014

    Figure Lengend Snippet: TREK2 knockdown in vivo decreased SFL 1 d after CFA. Ipsilateral SFL in rats 1 d after an intradermal plantar CFA injection in one hind foot in the same region as an intradermal plantar injection of either TREK2 siRNA or scrambled siRNA given 3 d earlier. Numbers in B and C (superscripts) and D and E (beside data points) relate to rats from which tissue in the lanes of A was obtained; these numbers are included to enable comparison of TREK2 protein expression with other data from the same rats. A , TREK2 Western blot of skin tissue, at the site of scr (1, 2) or siRNA (3, 4) injection 4 d previously, and 1 d after either saline (1, 3) or CFA injection (2, 4) at the same site. Top, The lower TREK2 band (MW ∼50 kDa, the band that we and others find in DRG tissues) was almost eliminated by siRNA, not scr, treatment, regardless of whether rats were injected with CFA or saline. The upper band was not reduced after siRNA. The MWs of these bands are consistent with them being two of the three known rat TREK2 isoforms (see Results and Discussion). Bottom, α-Tubulin loading controls. B , TREK2 knockdown by TREK2 siRNA in nerve fibers and epidermis (Ep) in plantar skin. Left, after scr siRNA injection. Right, after TREK2 siRNA injection. Both treatments were followed by CFA injection. Tissue was taken 4 d after scr/siRNA treatment (1 d after CFA). Triple fluorescence staining for β-tubulin III (blue, to stain nerve fibers), IB4 binding (green), and TREK2 (red). White dashed lines indicate junctions between epidermis (above) and dermis (below). In the merged images, colocalization of all three markers shows as white or very pale (bottom left image). After siRNA, red TREK2 staining in epidermis and in groups of nerve fibers (shown as blue and/or green) was reduced, showing substantial knockdown in vivo of TREK2 near the siRNA injection site. Scale bar, 25 μm. C , TREK2 knockdown by local TREK2 siRNA in the nerve innervating plantar skin. This knockdown, seen 5–10 mm from the siRNA injection site, occurred whether rats were subsequently treated with saline or CFA. There was decreased fiber TREK2 immunostaining (red) in TREK2 siRNA, not scr siRNA, treated rats, so that IB4 + fibers (green) showed little/no TREK2 (red). Scale bar, 50 μm. D , E , SFL after CFA increased after TREK2 siRNA compared with scr control. Ipsilateral SFLd ( D ) or SFLn ( E ) after siRNA C compared with scr treatment did not increase before saline injection (day 3, columns 1 and 2) or after saline injection (day 4, columns 3 and 4), or before CFA injection (day 3, columns 5 and 6). However, after CFA, it caused increased SFLd ( D ) and SFLn ( E ) in both scr ( p

    Article Snippet: Three days after transfection, the HEK cell cultures expressing the different K2P channels were washed with PBS, fixed with 4% paraformaldehyde in 4.2% sucrose (in PBS, pH 7.2) for 10 min at room temperature, rinsed with PBS, and processed for TREK2 immunocytochemistry with the rabbit anti-TREK2 antibody from Alomone Labs as described above.

    Techniques: In Vivo, Injection, Expressing, Western Blot, Fluorescence, Staining, Binding Assay, Immunostaining

    A , SF rate was related to Em in C-neurons in neuropathic pain models. SF rate in intact L4 C-nociceptors was significantly linearly correlated with Em 7 d after SNA and mSNA surgery (see Materials and Methods and Results). B , C , SFL 1 d after CFA was related to ipsilateral TREK2 levels. In ipsilateral L5 DRG small neurons in 5 rats 1 d after CFA-induced cutaneous inflammation, there were significant linear correlations of ( B ) SFLd and ( C ) number of SFLn with median ipsilateral L5 small DRG neuron TREK2 pixel densities for each rat, whether in entire cytoplasm, or edge, mid, or inner cytoplasmic regions. Thus, SFL after CFA in individual rats was related to ipsilateral TREK2 levels in small DRG neurons.

    Journal: The Journal of Neuroscience

    Article Title: TREK2 Expressed Selectively in IB4-Binding C-Fiber Nociceptors Hyperpolarizes Their Membrane Potentials and Limits Spontaneous Pain

    doi: 10.1523/JNEUROSCI.4528-13.2014

    Figure Lengend Snippet: A , SF rate was related to Em in C-neurons in neuropathic pain models. SF rate in intact L4 C-nociceptors was significantly linearly correlated with Em 7 d after SNA and mSNA surgery (see Materials and Methods and Results). B , C , SFL 1 d after CFA was related to ipsilateral TREK2 levels. In ipsilateral L5 DRG small neurons in 5 rats 1 d after CFA-induced cutaneous inflammation, there were significant linear correlations of ( B ) SFLd and ( C ) number of SFLn with median ipsilateral L5 small DRG neuron TREK2 pixel densities for each rat, whether in entire cytoplasm, or edge, mid, or inner cytoplasmic regions. Thus, SFL after CFA in individual rats was related to ipsilateral TREK2 levels in small DRG neurons.

    Article Snippet: Three days after transfection, the HEK cell cultures expressing the different K2P channels were washed with PBS, fixed with 4% paraformaldehyde in 4.2% sucrose (in PBS, pH 7.2) for 10 min at room temperature, rinsed with PBS, and processed for TREK2 immunocytochemistry with the rabbit anti-TREK2 antibody from Alomone Labs as described above.

    Techniques: