anti trpm3  (Alomone Labs)


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    Name:
    Anti TRPM3 extracellular Antibody
    Description:
    Anti TRPM3 extracellular Antibody ACC 050 is a highly specific antibody directed against an epitope of the human protein The antibody can be used in western blot immunocytochemistry and immunohistochemistry applications It has been designed to recognize TRPM3 from human rat and mouse samples
    Catalog Number:
    ACC-050
    Price:
    495.0
    Category:
    Primary Antibody
    Applications:
    Immunocytochemistry, Immunofluorescence, Immunohistochemistry, Western Blot
    Purity:
    Affinity purified on immobilized antigen.
    Immunogen:
    Synthetic peptide
    Size:
    50 mcl
    Antibody Type:
    Polyclonal Primary Antibodies
    Format:
    Lyophilized Powder
    Host:
    Rabbit
    Isotype:
    Rabbit IgG
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    Structured Review

    Alomone Labs anti trpm3
    Anti TRPM3 extracellular Antibody
    Anti TRPM3 extracellular Antibody ACC 050 is a highly specific antibody directed against an epitope of the human protein The antibody can be used in western blot immunocytochemistry and immunohistochemistry applications It has been designed to recognize TRPM3 from human rat and mouse samples
    https://www.bioz.com/result/anti trpm3/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti trpm3 - by Bioz Stars, 2021-09
    94/100 stars

    Images

    1) Product Images from "Store-operated calcium entry is essential for glial calcium signalling in CNS white matter"

    Article Title: Store-operated calcium entry is essential for glial calcium signalling in CNS white matter

    Journal: Brain Structure & Function

    doi: 10.1007/s00429-017-1380-8

    Mechanisms of SOCE in optic nerve glia. ATP-mediated calcium signalling in optic nerve glia is via P2Y G-protein-coupled receptors and the formation of IP3, which acts on IP3R1 on the ER to trigger release of Ca 2+ into the cytosol. Subsequent replenishment of ER stores in astrocytes and oligodendrocytes is dependent on SOCE via TRPM3 and Orai1, which form the plasmalemmal Ca 2+ channels, and mainly Stim1, which acts as the sensor of Ca 2+ depletion, and uptake into the ER is via SERCA pumps. Oligodendrocytes also express Stim2, which may be localized to the myelin, whereas Orai1, Stim1 and TRPM3 are localized to oligodendroglial somata. Notably, calcium homeostasis in optic nerve glia depends on an apparent continuous Ca 2+ influx from the extracellular milieu that is largely dependent on SOCE. Moreover, SOCE is essential for the sustainability of ATP-mediated Ca 2+ signalling in optic nerve glia, which has a central role in white matter physiology and pathology
    Figure Legend Snippet: Mechanisms of SOCE in optic nerve glia. ATP-mediated calcium signalling in optic nerve glia is via P2Y G-protein-coupled receptors and the formation of IP3, which acts on IP3R1 on the ER to trigger release of Ca 2+ into the cytosol. Subsequent replenishment of ER stores in astrocytes and oligodendrocytes is dependent on SOCE via TRPM3 and Orai1, which form the plasmalemmal Ca 2+ channels, and mainly Stim1, which acts as the sensor of Ca 2+ depletion, and uptake into the ER is via SERCA pumps. Oligodendrocytes also express Stim2, which may be localized to the myelin, whereas Orai1, Stim1 and TRPM3 are localized to oligodendroglial somata. Notably, calcium homeostasis in optic nerve glia depends on an apparent continuous Ca 2+ influx from the extracellular milieu that is largely dependent on SOCE. Moreover, SOCE is essential for the sustainability of ATP-mediated Ca 2+ signalling in optic nerve glia, which has a central role in white matter physiology and pathology

    Techniques Used:

    Expression of TRP channels in optic nerve glia. a qRT-PCR of acutely isolated optic nerves from WT mice aged P9–P12 and P30–P40; data are from 10 pooled optic nerves in each age group, run in triplicate, expressed as relative mRNA levels (2 -ΔCt ) compared to the housekeeping gene GAPDH method (mean ± SEM, n = 3). TRPM3 was the most highly expressed TRP channel in the postnatal and adult nerve (*** p
    Figure Legend Snippet: Expression of TRP channels in optic nerve glia. a qRT-PCR of acutely isolated optic nerves from WT mice aged P9–P12 and P30–P40; data are from 10 pooled optic nerves in each age group, run in triplicate, expressed as relative mRNA levels (2 -ΔCt ) compared to the housekeeping gene GAPDH method (mean ± SEM, n = 3). TRPM3 was the most highly expressed TRP channel in the postnatal and adult nerve (*** p

    Techniques Used: Expressing, Quantitative RT-PCR, Isolation, Mouse Assay

    2) Product Images from "The effect of IL-2 stimulation and treatment of TRPM3 on channel co-localisation with PIP2 and NK cell function in myalgic encephalomyelitis/chronic fatigue syndrome patients"

    Article Title: The effect of IL-2 stimulation and treatment of TRPM3 on channel co-localisation with PIP2 and NK cell function in myalgic encephalomyelitis/chronic fatigue syndrome patients

    Journal: Journal of Translational Medicine

    doi: 10.1186/s12967-021-02974-4

    A Mann Whitney U test using PCC and MOC co-localisation values of TRPM3 with actin on NK cells of ME/CFS patients and HC. B Mann Whitney U test using PCC and MOC co-localisation values of TRPM3 with PIP 2 on NK cells of ME/CFS patients and HC. Co-localisation values obtained following IL-2 stimulation (20 IU) (control) and pharmacological treatment using PregS (30 µM) and PregS + Ononetin (3 µM). Bar graphs represent correlation between target antigens using PCC represent degree of overlap between target antigens using MOC. Data presented as mean ± SEM and *p
    Figure Legend Snippet: A Mann Whitney U test using PCC and MOC co-localisation values of TRPM3 with actin on NK cells of ME/CFS patients and HC. B Mann Whitney U test using PCC and MOC co-localisation values of TRPM3 with PIP 2 on NK cells of ME/CFS patients and HC. Co-localisation values obtained following IL-2 stimulation (20 IU) (control) and pharmacological treatment using PregS (30 µM) and PregS + Ononetin (3 µM). Bar graphs represent correlation between target antigens using PCC represent degree of overlap between target antigens using MOC. Data presented as mean ± SEM and *p

    Techniques Used: MANN-WHITNEY, Periodic Counter-current Chromatography

    Co-localisation and immunofluorescent images of TRPM3 with actin in NK cells of HC. A Example of immunostaining of actin (phalloidin, red), nucleus (DAPI, blue), TRPM3 (green) in an NKLa cell under control IL-2 conditions. Cells were stimulated overnight with IL-2 (20 IU) (control) and treated with PregS (30 µM) and ononetin (3 µM). Images taken using Nikon A1R + confocal microscopy. B Bar graphs represent correlation between target antigens using Pearson’s correlation coefficient and the degree of overlap between target antigens using Mander’s overlap coefficient. Co-localisation coefficients K1 (TRPM3) and K2 (actin) were used to determine contribution to co-localisation. Number of cells analysed are included within bar graphs: Control n = 260; PregS n = 277; and PregS + Ono n = 219. Data presented as mean ± SEM and *p
    Figure Legend Snippet: Co-localisation and immunofluorescent images of TRPM3 with actin in NK cells of HC. A Example of immunostaining of actin (phalloidin, red), nucleus (DAPI, blue), TRPM3 (green) in an NKLa cell under control IL-2 conditions. Cells were stimulated overnight with IL-2 (20 IU) (control) and treated with PregS (30 µM) and ononetin (3 µM). Images taken using Nikon A1R + confocal microscopy. B Bar graphs represent correlation between target antigens using Pearson’s correlation coefficient and the degree of overlap between target antigens using Mander’s overlap coefficient. Co-localisation coefficients K1 (TRPM3) and K2 (actin) were used to determine contribution to co-localisation. Number of cells analysed are included within bar graphs: Control n = 260; PregS n = 277; and PregS + Ono n = 219. Data presented as mean ± SEM and *p

    Techniques Used: Immunostaining, Confocal Microscopy

    Co-localisation and immunofluorescent images of TRPM3 with actin in NK cells of ME/CFS patients. A Example of immunostaining of actin (phalloidin, red), nucleus (DAPI, blue), TRPM3 (green) in NK cell under control IL-2 condition. Cells were stimulated overnight with IL-2 (20 IU) (control) and treated with PregS (30 µM) and ononetin (3 µM). Images taken using Nikon A1R + confocal microscopy. B Bar graphs represent correlation between target antigens using Pearson’s correlation coefficient and the degree of overlap between target antigens using Mander’s overlap coefficient. Co-localisation coefficients K1 (TRPM3) and K2 (actin) were used to determine contribution to co-localisation. Number of cells analysed are included within bar graphs: Control n = 185; PregS n = 193; and PregS + Ono n = 197. Data presented as mean ± SEM. TRPM3, transient receptor potential Melastatin; PregS, pregnenolone sulfate; Ono, ononetin; IL-2, interleukin 2
    Figure Legend Snippet: Co-localisation and immunofluorescent images of TRPM3 with actin in NK cells of ME/CFS patients. A Example of immunostaining of actin (phalloidin, red), nucleus (DAPI, blue), TRPM3 (green) in NK cell under control IL-2 condition. Cells were stimulated overnight with IL-2 (20 IU) (control) and treated with PregS (30 µM) and ononetin (3 µM). Images taken using Nikon A1R + confocal microscopy. B Bar graphs represent correlation between target antigens using Pearson’s correlation coefficient and the degree of overlap between target antigens using Mander’s overlap coefficient. Co-localisation coefficients K1 (TRPM3) and K2 (actin) were used to determine contribution to co-localisation. Number of cells analysed are included within bar graphs: Control n = 185; PregS n = 193; and PregS + Ono n = 197. Data presented as mean ± SEM. TRPM3, transient receptor potential Melastatin; PregS, pregnenolone sulfate; Ono, ononetin; IL-2, interleukin 2

    Techniques Used: Immunostaining, Confocal Microscopy

    Co-localisation and immunofluorescent images of TRPM3 with PIP 2 in NK cells of ME/CFS. patients. A Example of immunostaining of PIP 2 (red), nucleus (DAPI, blue), TRPM3 (green) in NK cell under control IL-2 conditions. Cells were stimulated overnight with IL-2 (20 IU) (control) and treated with PregS (30 µM) and ononetin (3 µM). Images taken using Nikon A1R + confocal microscopy. B Bar graphs represent correlation between target antigens using Pearson’s correlation coefficient and the degree of overlap between target antigens using Mander’s overlap coefficient. Co-localisation coefficients K1 (TRPM3) and K2 (PIP 2 ) were used to determine contribution to co-localisation. Number of cells analysed are included within bar graphs: Control n = 214; PregS n = 225; and PregS + Ono n = 220. Data presented as mean ± SEM and *p
    Figure Legend Snippet: Co-localisation and immunofluorescent images of TRPM3 with PIP 2 in NK cells of ME/CFS. patients. A Example of immunostaining of PIP 2 (red), nucleus (DAPI, blue), TRPM3 (green) in NK cell under control IL-2 conditions. Cells were stimulated overnight with IL-2 (20 IU) (control) and treated with PregS (30 µM) and ononetin (3 µM). Images taken using Nikon A1R + confocal microscopy. B Bar graphs represent correlation between target antigens using Pearson’s correlation coefficient and the degree of overlap between target antigens using Mander’s overlap coefficient. Co-localisation coefficients K1 (TRPM3) and K2 (PIP 2 ) were used to determine contribution to co-localisation. Number of cells analysed are included within bar graphs: Control n = 214; PregS n = 225; and PregS + Ono n = 220. Data presented as mean ± SEM and *p

    Techniques Used: Immunostaining, Confocal Microscopy

    Co-localisation and immunofluorescent images of TRPM3 with PIP 2 in NK cells of HC. A Example of immunostaining of PIP 2 (red), nucleus (DAPI, blue), TRPM3 (green)) in an NK cell under control IL-2 conditions. Images taken using Nikon A1R + confocal microscopy. B Bar graphs represent correlation between target antigens using Pearson’s correlation coefficient and the degree of overlap between target antigens using Mander’s overlap coefficient. Co-localisation coefficients K1 (TRPM3) and K2 (PIP 2 ) were used to determine contribution to co-localisation. Number of cells analysed are included within bar graphs: Control n = 287; PregS n = 292; and PregS + Ono n = 237. Data presented as mean ± SEM and *p
    Figure Legend Snippet: Co-localisation and immunofluorescent images of TRPM3 with PIP 2 in NK cells of HC. A Example of immunostaining of PIP 2 (red), nucleus (DAPI, blue), TRPM3 (green)) in an NK cell under control IL-2 conditions. Images taken using Nikon A1R + confocal microscopy. B Bar graphs represent correlation between target antigens using Pearson’s correlation coefficient and the degree of overlap between target antigens using Mander’s overlap coefficient. Co-localisation coefficients K1 (TRPM3) and K2 (PIP 2 ) were used to determine contribution to co-localisation. Number of cells analysed are included within bar graphs: Control n = 287; PregS n = 292; and PregS + Ono n = 237. Data presented as mean ± SEM and *p

    Techniques Used: Immunostaining, Confocal Microscopy

    3) Product Images from "Functional expression and pharmacological modulation of TRPM3 in human sensory neurons, et al. Functional expression and pharmacological modulation of TRPM3 in human sensory neurons"

    Article Title: Functional expression and pharmacological modulation of TRPM3 in human sensory neurons, et al. Functional expression and pharmacological modulation of TRPM3 in human sensory neurons

    Journal: British Journal of Pharmacology

    doi: 10.1111/bph.14994

    Electrophysiological properties of TRPM3 in human stem cell‐derived sensory neurons (hSCDS) neurons. (a) Time course of inward (at −120 mV) and outward (at +80 mV) whole‐cell currents in hSCDS neurons showing the effects of the TRPM3 agonists pregnenolone sulphate (PS; 40 μM) and CIM0216 (1 μM). (b) Quantification of the amplitude of inward and outward currents activated by PS and CIM0216 ( n = ]
    Figure Legend Snippet: Electrophysiological properties of TRPM3 in human stem cell‐derived sensory neurons (hSCDS) neurons. (a) Time course of inward (at −120 mV) and outward (at +80 mV) whole‐cell currents in hSCDS neurons showing the effects of the TRPM3 agonists pregnenolone sulphate (PS; 40 μM) and CIM0216 (1 μM). (b) Quantification of the amplitude of inward and outward currents activated by PS and CIM0216 ( n = ]

    Techniques Used: Derivative Assay

    Pharmacological characterization of TRPM3 in human stem cell‐derived sensory neurons (hSCDS) neurons. (a) Time course of the intracellular Ca 2+ concentration in PS responding hSCDS neurons upon increasing the concentration of pregnenolone sulphate (PS) from 1 to 300 μM. (b) Concentration dependence of the PS‐induced calcium response in hSCDS neurons. Responses were normalized to the maximal response ( n = 117). Solid line represents a fit using a Hill function. (c) Normalized calcium traces showing the effect of increasing concentrations of the TRPM3 antagonist isosakuranetin (Isoas) on the response to PS (40 μM). Green trace represents the vehicle control, showing mild rundown of the signal in the absence of antagonist. (d) Concentration–response curve for the inhibition of PS‐evoked calcium responses by isosakuranetin ( n = 93; red squares) and primidone ( n = ]
    Figure Legend Snippet: Pharmacological characterization of TRPM3 in human stem cell‐derived sensory neurons (hSCDS) neurons. (a) Time course of the intracellular Ca 2+ concentration in PS responding hSCDS neurons upon increasing the concentration of pregnenolone sulphate (PS) from 1 to 300 μM. (b) Concentration dependence of the PS‐induced calcium response in hSCDS neurons. Responses were normalized to the maximal response ( n = 117). Solid line represents a fit using a Hill function. (c) Normalized calcium traces showing the effect of increasing concentrations of the TRPM3 antagonist isosakuranetin (Isoas) on the response to PS (40 μM). Green trace represents the vehicle control, showing mild rundown of the signal in the absence of antagonist. (d) Concentration–response curve for the inhibition of PS‐evoked calcium responses by isosakuranetin ( n = 93; red squares) and primidone ( n = ]

    Techniques Used: Derivative Assay, Concentration Assay, Inhibition

    Functional TRPM3 expression in human dorsal root ganglion (DRG) neurons. (a) Expression levels of TRP channel mRNA relative to HPRT in DRG neurons from two donors, determined using RT‐qPCR. Access to fresh human DRG tissue is scarce, hence the limited number of samples in this exploratory experiment. nd, not detected. (b) Representative example of changes in Fluo‐8‐fluorescence in human DRG neurons in response to the TRPM3 agonists pregnenolone sulphate (PS; 50 μM) and CIM0216 (10 μM) and to the TRPV1 agonist capsaicin (Caps; 200 nM). A high K + . (c) Pie chart showing the distribution of neurons responding to PS, capsaicin or both ( n . (e) Normalized responses to repeated PS applications. Neurons were stimulated three times with PS, and the second application occurred in the presence of either isosakuranetin (10 μM; n = 50) or vehicle control ( n = 39). * P ]
    Figure Legend Snippet: Functional TRPM3 expression in human dorsal root ganglion (DRG) neurons. (a) Expression levels of TRP channel mRNA relative to HPRT in DRG neurons from two donors, determined using RT‐qPCR. Access to fresh human DRG tissue is scarce, hence the limited number of samples in this exploratory experiment. nd, not detected. (b) Representative example of changes in Fluo‐8‐fluorescence in human DRG neurons in response to the TRPM3 agonists pregnenolone sulphate (PS; 50 μM) and CIM0216 (10 μM) and to the TRPV1 agonist capsaicin (Caps; 200 nM). A high K + . (c) Pie chart showing the distribution of neurons responding to PS, capsaicin or both ( n . (e) Normalized responses to repeated PS applications. Neurons were stimulated three times with PS, and the second application occurred in the presence of either isosakuranetin (10 μM; n = 50) or vehicle control ( n = 39). * P ]

    Techniques Used: Functional Assay, Expressing, Quantitative RT-PCR, Fluorescence

    Modulation of TRPM3 in human stem cell‐derived sensory neurons (hSCDS) neurons by GPCRs. (a) Example intracellular calcium measurements showing variable degrees of inhibition of PS‐induced responses upon activation of endogenously expressed μ‐opioid receptors using DAMGO (300 nM). (b) Scatter plot showing the range of DAMGO‐induced inhibition of PS responses in hSCDS neurons. Red line represents the mean percentage of TRPM3 inhibition caused by DAMGO (data from six experiments, with hSCDS neurons from three different differentiations). (c) Example intracellular calcium traces showing inhibition of PS‐induced responses by the GABA B ]
    Figure Legend Snippet: Modulation of TRPM3 in human stem cell‐derived sensory neurons (hSCDS) neurons by GPCRs. (a) Example intracellular calcium measurements showing variable degrees of inhibition of PS‐induced responses upon activation of endogenously expressed μ‐opioid receptors using DAMGO (300 nM). (b) Scatter plot showing the range of DAMGO‐induced inhibition of PS responses in hSCDS neurons. Red line represents the mean percentage of TRPM3 inhibition caused by DAMGO (data from six experiments, with hSCDS neurons from three different differentiations). (c) Example intracellular calcium traces showing inhibition of PS‐induced responses by the GABA B ]

    Techniques Used: Derivative Assay, Inhibition, Activation Assay

    Functional expression profile of somatosensory TRP channels in human stem cell‐derived sensory neurons (hSCDS) neurons. (a) Changes in intracellular calcium in single hSCDS neurons stimulated with the TRP channel agonists menthol (100 μM), MO (100 μM), capsaicin (Caps;1 μM), and pregnenolone sulphate (PS;40 μM) and with a high K + solution to probe excitability. (b) Venn diagram showing the pattern of responses to TRP channel agonists in hSCDS neurons ( n = 1,180 cells in five independent differentiations). (c) Intracellular calcium measurements showing reversible inhibition of PS‐evoked responses by isosakuranetin (5 μM). PS‐responsive hSCDS neurons also responded to the synthetic TRPM3 agonist CIM0216 (1 μM). (d) Quantification of calcium responses for experiments as in panel (c) ( n = ]
    Figure Legend Snippet: Functional expression profile of somatosensory TRP channels in human stem cell‐derived sensory neurons (hSCDS) neurons. (a) Changes in intracellular calcium in single hSCDS neurons stimulated with the TRP channel agonists menthol (100 μM), MO (100 μM), capsaicin (Caps;1 μM), and pregnenolone sulphate (PS;40 μM) and with a high K + solution to probe excitability. (b) Venn diagram showing the pattern of responses to TRP channel agonists in hSCDS neurons ( n = 1,180 cells in five independent differentiations). (c) Intracellular calcium measurements showing reversible inhibition of PS‐evoked responses by isosakuranetin (5 μM). PS‐responsive hSCDS neurons also responded to the synthetic TRPM3 agonist CIM0216 (1 μM). (d) Quantification of calcium responses for experiments as in panel (c) ( n = ]

    Techniques Used: Functional Assay, Expressing, Derivative Assay, Inhibition

    4) Product Images from "The effect of IL-2 stimulation and treatment of TRPM3 on channel co-localisation with PIP2 and NK cell function in myalgic encephalomyelitis/chronic fatigue syndrome patients"

    Article Title: The effect of IL-2 stimulation and treatment of TRPM3 on channel co-localisation with PIP2 and NK cell function in myalgic encephalomyelitis/chronic fatigue syndrome patients

    Journal: Journal of Translational Medicine

    doi: 10.1186/s12967-021-02974-4

    A Mann Whitney U test using PCC and MOC co-localisation values of TRPM3 with actin on NK cells of ME/CFS patients and HC. B Mann Whitney U test using PCC and MOC co-localisation values of TRPM3 with PIP 2 on NK cells of ME/CFS patients and HC. Co-localisation values obtained following IL-2 stimulation (20 IU) (control) and pharmacological treatment using PregS (30 µM) and PregS + Ononetin (3 µM). Bar graphs represent correlation between target antigens using PCC represent degree of overlap between target antigens using MOC. Data presented as mean ± SEM and *p
    Figure Legend Snippet: A Mann Whitney U test using PCC and MOC co-localisation values of TRPM3 with actin on NK cells of ME/CFS patients and HC. B Mann Whitney U test using PCC and MOC co-localisation values of TRPM3 with PIP 2 on NK cells of ME/CFS patients and HC. Co-localisation values obtained following IL-2 stimulation (20 IU) (control) and pharmacological treatment using PregS (30 µM) and PregS + Ononetin (3 µM). Bar graphs represent correlation between target antigens using PCC represent degree of overlap between target antigens using MOC. Data presented as mean ± SEM and *p

    Techniques Used: MANN-WHITNEY, Periodic Counter-current Chromatography

    Co-localisation and immunofluorescent images of TRPM3 with actin in NK cells of HC. A Example of immunostaining of actin (phalloidin, red), nucleus (DAPI, blue), TRPM3 (green) in an NKLa cell under control IL-2 conditions. Cells were stimulated overnight with IL-2 (20 IU) (control) and treated with PregS (30 µM) and ononetin (3 µM). Images taken using Nikon A1R + confocal microscopy. B Bar graphs represent correlation between target antigens using Pearson’s correlation coefficient and the degree of overlap between target antigens using Mander’s overlap coefficient. Co-localisation coefficients K1 (TRPM3) and K2 (actin) were used to determine contribution to co-localisation. Number of cells analysed are included within bar graphs: Control n = 260; PregS n = 277; and PregS + Ono n = 219. Data presented as mean ± SEM and *p
    Figure Legend Snippet: Co-localisation and immunofluorescent images of TRPM3 with actin in NK cells of HC. A Example of immunostaining of actin (phalloidin, red), nucleus (DAPI, blue), TRPM3 (green) in an NKLa cell under control IL-2 conditions. Cells were stimulated overnight with IL-2 (20 IU) (control) and treated with PregS (30 µM) and ononetin (3 µM). Images taken using Nikon A1R + confocal microscopy. B Bar graphs represent correlation between target antigens using Pearson’s correlation coefficient and the degree of overlap between target antigens using Mander’s overlap coefficient. Co-localisation coefficients K1 (TRPM3) and K2 (actin) were used to determine contribution to co-localisation. Number of cells analysed are included within bar graphs: Control n = 260; PregS n = 277; and PregS + Ono n = 219. Data presented as mean ± SEM and *p

    Techniques Used: Immunostaining, Confocal Microscopy

    Co-localisation and immunofluorescent images of TRPM3 with actin in NK cells of ME/CFS patients. A Example of immunostaining of actin (phalloidin, red), nucleus (DAPI, blue), TRPM3 (green) in NK cell under control IL-2 condition. Cells were stimulated overnight with IL-2 (20 IU) (control) and treated with PregS (30 µM) and ononetin (3 µM). Images taken using Nikon A1R + confocal microscopy. B Bar graphs represent correlation between target antigens using Pearson’s correlation coefficient and the degree of overlap between target antigens using Mander’s overlap coefficient. Co-localisation coefficients K1 (TRPM3) and K2 (actin) were used to determine contribution to co-localisation. Number of cells analysed are included within bar graphs: Control n = 185; PregS n = 193; and PregS + Ono n = 197. Data presented as mean ± SEM. TRPM3, transient receptor potential Melastatin; PregS, pregnenolone sulfate; Ono, ononetin; IL-2, interleukin 2
    Figure Legend Snippet: Co-localisation and immunofluorescent images of TRPM3 with actin in NK cells of ME/CFS patients. A Example of immunostaining of actin (phalloidin, red), nucleus (DAPI, blue), TRPM3 (green) in NK cell under control IL-2 condition. Cells were stimulated overnight with IL-2 (20 IU) (control) and treated with PregS (30 µM) and ononetin (3 µM). Images taken using Nikon A1R + confocal microscopy. B Bar graphs represent correlation between target antigens using Pearson’s correlation coefficient and the degree of overlap between target antigens using Mander’s overlap coefficient. Co-localisation coefficients K1 (TRPM3) and K2 (actin) were used to determine contribution to co-localisation. Number of cells analysed are included within bar graphs: Control n = 185; PregS n = 193; and PregS + Ono n = 197. Data presented as mean ± SEM. TRPM3, transient receptor potential Melastatin; PregS, pregnenolone sulfate; Ono, ononetin; IL-2, interleukin 2

    Techniques Used: Immunostaining, Confocal Microscopy

    Co-localisation and immunofluorescent images of TRPM3 with PIP 2 in NK cells of ME/CFS. patients. A Example of immunostaining of PIP 2 (red), nucleus (DAPI, blue), TRPM3 (green) in NK cell under control IL-2 conditions. Cells were stimulated overnight with IL-2 (20 IU) (control) and treated with PregS (30 µM) and ononetin (3 µM). Images taken using Nikon A1R + confocal microscopy. B Bar graphs represent correlation between target antigens using Pearson’s correlation coefficient and the degree of overlap between target antigens using Mander’s overlap coefficient. Co-localisation coefficients K1 (TRPM3) and K2 (PIP 2 ) were used to determine contribution to co-localisation. Number of cells analysed are included within bar graphs: Control n = 214; PregS n = 225; and PregS + Ono n = 220. Data presented as mean ± SEM and *p
    Figure Legend Snippet: Co-localisation and immunofluorescent images of TRPM3 with PIP 2 in NK cells of ME/CFS. patients. A Example of immunostaining of PIP 2 (red), nucleus (DAPI, blue), TRPM3 (green) in NK cell under control IL-2 conditions. Cells were stimulated overnight with IL-2 (20 IU) (control) and treated with PregS (30 µM) and ononetin (3 µM). Images taken using Nikon A1R + confocal microscopy. B Bar graphs represent correlation between target antigens using Pearson’s correlation coefficient and the degree of overlap between target antigens using Mander’s overlap coefficient. Co-localisation coefficients K1 (TRPM3) and K2 (PIP 2 ) were used to determine contribution to co-localisation. Number of cells analysed are included within bar graphs: Control n = 214; PregS n = 225; and PregS + Ono n = 220. Data presented as mean ± SEM and *p

    Techniques Used: Immunostaining, Confocal Microscopy

    Co-localisation and immunofluorescent images of TRPM3 with PIP 2 in NK cells of HC. A Example of immunostaining of PIP 2 (red), nucleus (DAPI, blue), TRPM3 (green)) in an NK cell under control IL-2 conditions. Images taken using Nikon A1R + confocal microscopy. B Bar graphs represent correlation between target antigens using Pearson’s correlation coefficient and the degree of overlap between target antigens using Mander’s overlap coefficient. Co-localisation coefficients K1 (TRPM3) and K2 (PIP 2 ) were used to determine contribution to co-localisation. Number of cells analysed are included within bar graphs: Control n = 287; PregS n = 292; and PregS + Ono n = 237. Data presented as mean ± SEM and *p
    Figure Legend Snippet: Co-localisation and immunofluorescent images of TRPM3 with PIP 2 in NK cells of HC. A Example of immunostaining of PIP 2 (red), nucleus (DAPI, blue), TRPM3 (green)) in an NK cell under control IL-2 conditions. Images taken using Nikon A1R + confocal microscopy. B Bar graphs represent correlation between target antigens using Pearson’s correlation coefficient and the degree of overlap between target antigens using Mander’s overlap coefficient. Co-localisation coefficients K1 (TRPM3) and K2 (PIP 2 ) were used to determine contribution to co-localisation. Number of cells analysed are included within bar graphs: Control n = 287; PregS n = 292; and PregS + Ono n = 237. Data presented as mean ± SEM and *p

    Techniques Used: Immunostaining, Confocal Microscopy

    Related Articles

    Incubation:

    Article Title: Transient Receptor Potential Melastatin-3 (TRPM3) Mediates Nociceptive-Like Responses in Hydra vulgaris
    Article Snippet: .. Membrane was incubated with rabbit polyclonal anti-TRPM3 antibody (O/N, 4°C, 1:100 dilution; Alomone Labs, Jerusalem, Israel). ..

    Article Title: Store-operated calcium entry is essential for glial calcium signalling in CNS white matter
    Article Snippet: .. Primary antibodies were diluted in blocking solution and tissues/cells incubated overnight at 4 °C; anti-STIM1, anti-STIM2, anti-ORAI1, anti-TRPM3 were raised in rabbits (Alomone) and used at 1:300; chicken anti-GFAP (Chemicon) was used at 1:500. ..

    Article Title: The effect of IL-2 stimulation and treatment of TRPM3 on channel co-localisation with PIP2 and NK cell function in myalgic encephalomyelitis/chronic fatigue syndrome patients
    Article Snippet: .. NK cells were incubated overnight (16 h) at 4 °C with primary antibodies for TRPM3 (1:6,000) (Alomone, Jerusalem, Israel) and PIP2 (1:3,000) (Abcam, Cambridge, UK) in PBS 1X + 3% BSA. ..

    Article Title: Functional expression and pharmacological modulation of TRPM3 in human sensory neurons, et al. Functional expression and pharmacological modulation of TRPM3 in human sensory neurons
    Article Snippet: .. After blocking, TRPM3 antibody (Alomone Labs, Cat# ACC‐050, RRID:AB_10918820; 0.2 μg·ml−1 ) was incubated overnight. ..

    Blocking Assay:

    Article Title: Store-operated calcium entry is essential for glial calcium signalling in CNS white matter
    Article Snippet: .. Primary antibodies were diluted in blocking solution and tissues/cells incubated overnight at 4 °C; anti-STIM1, anti-STIM2, anti-ORAI1, anti-TRPM3 were raised in rabbits (Alomone) and used at 1:300; chicken anti-GFAP (Chemicon) was used at 1:500. ..

    Article Title: Functional expression and pharmacological modulation of TRPM3 in human sensory neurons, et al. Functional expression and pharmacological modulation of TRPM3 in human sensory neurons
    Article Snippet: .. After blocking, TRPM3 antibody (Alomone Labs, Cat# ACC‐050, RRID:AB_10918820; 0.2 μg·ml−1 ) was incubated overnight. ..

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    Alomone Labs anti trpm3
    Mechanisms of SOCE in optic nerve glia. ATP-mediated calcium signalling in optic nerve glia is via P2Y G-protein-coupled receptors and the formation of IP3, which acts on IP3R1 on the ER to trigger release of Ca 2+ into the cytosol. Subsequent replenishment of ER stores in astrocytes and oligodendrocytes is dependent on SOCE via <t>TRPM3</t> and Orai1, which form the plasmalemmal Ca 2+ channels, and mainly Stim1, which acts as the sensor of Ca 2+ depletion, and uptake into the ER is via SERCA pumps. Oligodendrocytes also express Stim2, which may be localized to the myelin, whereas Orai1, Stim1 and TRPM3 are localized to oligodendroglial somata. Notably, calcium homeostasis in optic nerve glia depends on an apparent continuous Ca 2+ influx from the extracellular milieu that is largely dependent on SOCE. Moreover, SOCE is essential for the sustainability of ATP-mediated Ca 2+ signalling in optic nerve glia, which has a central role in white matter physiology and pathology
    Anti Trpm3, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Mechanisms of SOCE in optic nerve glia. ATP-mediated calcium signalling in optic nerve glia is via P2Y G-protein-coupled receptors and the formation of IP3, which acts on IP3R1 on the ER to trigger release of Ca 2+ into the cytosol. Subsequent replenishment of ER stores in astrocytes and oligodendrocytes is dependent on SOCE via TRPM3 and Orai1, which form the plasmalemmal Ca 2+ channels, and mainly Stim1, which acts as the sensor of Ca 2+ depletion, and uptake into the ER is via SERCA pumps. Oligodendrocytes also express Stim2, which may be localized to the myelin, whereas Orai1, Stim1 and TRPM3 are localized to oligodendroglial somata. Notably, calcium homeostasis in optic nerve glia depends on an apparent continuous Ca 2+ influx from the extracellular milieu that is largely dependent on SOCE. Moreover, SOCE is essential for the sustainability of ATP-mediated Ca 2+ signalling in optic nerve glia, which has a central role in white matter physiology and pathology

    Journal: Brain Structure & Function

    Article Title: Store-operated calcium entry is essential for glial calcium signalling in CNS white matter

    doi: 10.1007/s00429-017-1380-8

    Figure Lengend Snippet: Mechanisms of SOCE in optic nerve glia. ATP-mediated calcium signalling in optic nerve glia is via P2Y G-protein-coupled receptors and the formation of IP3, which acts on IP3R1 on the ER to trigger release of Ca 2+ into the cytosol. Subsequent replenishment of ER stores in astrocytes and oligodendrocytes is dependent on SOCE via TRPM3 and Orai1, which form the plasmalemmal Ca 2+ channels, and mainly Stim1, which acts as the sensor of Ca 2+ depletion, and uptake into the ER is via SERCA pumps. Oligodendrocytes also express Stim2, which may be localized to the myelin, whereas Orai1, Stim1 and TRPM3 are localized to oligodendroglial somata. Notably, calcium homeostasis in optic nerve glia depends on an apparent continuous Ca 2+ influx from the extracellular milieu that is largely dependent on SOCE. Moreover, SOCE is essential for the sustainability of ATP-mediated Ca 2+ signalling in optic nerve glia, which has a central role in white matter physiology and pathology

    Article Snippet: Primary antibodies were diluted in blocking solution and tissues/cells incubated overnight at 4 °C; anti-STIM1, anti-STIM2, anti-ORAI1, anti-TRPM3 were raised in rabbits (Alomone) and used at 1:300; chicken anti-GFAP (Chemicon) was used at 1:500.

    Techniques:

    Expression of TRP channels in optic nerve glia. a qRT-PCR of acutely isolated optic nerves from WT mice aged P9–P12 and P30–P40; data are from 10 pooled optic nerves in each age group, run in triplicate, expressed as relative mRNA levels (2 -ΔCt ) compared to the housekeeping gene GAPDH method (mean ± SEM, n = 3). TRPM3 was the most highly expressed TRP channel in the postnatal and adult nerve (*** p

    Journal: Brain Structure & Function

    Article Title: Store-operated calcium entry is essential for glial calcium signalling in CNS white matter

    doi: 10.1007/s00429-017-1380-8

    Figure Lengend Snippet: Expression of TRP channels in optic nerve glia. a qRT-PCR of acutely isolated optic nerves from WT mice aged P9–P12 and P30–P40; data are from 10 pooled optic nerves in each age group, run in triplicate, expressed as relative mRNA levels (2 -ΔCt ) compared to the housekeeping gene GAPDH method (mean ± SEM, n = 3). TRPM3 was the most highly expressed TRP channel in the postnatal and adult nerve (*** p

    Article Snippet: Primary antibodies were diluted in blocking solution and tissues/cells incubated overnight at 4 °C; anti-STIM1, anti-STIM2, anti-ORAI1, anti-TRPM3 were raised in rabbits (Alomone) and used at 1:300; chicken anti-GFAP (Chemicon) was used at 1:500.

    Techniques: Expressing, Quantitative RT-PCR, Isolation, Mouse Assay

    A Mann Whitney U test using PCC and MOC co-localisation values of TRPM3 with actin on NK cells of ME/CFS patients and HC. B Mann Whitney U test using PCC and MOC co-localisation values of TRPM3 with PIP 2 on NK cells of ME/CFS patients and HC. Co-localisation values obtained following IL-2 stimulation (20 IU) (control) and pharmacological treatment using PregS (30 µM) and PregS + Ononetin (3 µM). Bar graphs represent correlation between target antigens using PCC represent degree of overlap between target antigens using MOC. Data presented as mean ± SEM and *p

    Journal: Journal of Translational Medicine

    Article Title: The effect of IL-2 stimulation and treatment of TRPM3 on channel co-localisation with PIP2 and NK cell function in myalgic encephalomyelitis/chronic fatigue syndrome patients

    doi: 10.1186/s12967-021-02974-4

    Figure Lengend Snippet: A Mann Whitney U test using PCC and MOC co-localisation values of TRPM3 with actin on NK cells of ME/CFS patients and HC. B Mann Whitney U test using PCC and MOC co-localisation values of TRPM3 with PIP 2 on NK cells of ME/CFS patients and HC. Co-localisation values obtained following IL-2 stimulation (20 IU) (control) and pharmacological treatment using PregS (30 µM) and PregS + Ononetin (3 µM). Bar graphs represent correlation between target antigens using PCC represent degree of overlap between target antigens using MOC. Data presented as mean ± SEM and *p

    Article Snippet: Anti-PIP2 was purchased from Abcam (product code: ab11039) while anti-TRPM3 was purchased from Alomone Labs (Product code: ACC-050) and reconstituted at 0.8 mg/ml in distilled water.

    Techniques: MANN-WHITNEY, Periodic Counter-current Chromatography

    Co-localisation and immunofluorescent images of TRPM3 with actin in NK cells of HC. A Example of immunostaining of actin (phalloidin, red), nucleus (DAPI, blue), TRPM3 (green) in an NKLa cell under control IL-2 conditions. Cells were stimulated overnight with IL-2 (20 IU) (control) and treated with PregS (30 µM) and ononetin (3 µM). Images taken using Nikon A1R + confocal microscopy. B Bar graphs represent correlation between target antigens using Pearson’s correlation coefficient and the degree of overlap between target antigens using Mander’s overlap coefficient. Co-localisation coefficients K1 (TRPM3) and K2 (actin) were used to determine contribution to co-localisation. Number of cells analysed are included within bar graphs: Control n = 260; PregS n = 277; and PregS + Ono n = 219. Data presented as mean ± SEM and *p

    Journal: Journal of Translational Medicine

    Article Title: The effect of IL-2 stimulation and treatment of TRPM3 on channel co-localisation with PIP2 and NK cell function in myalgic encephalomyelitis/chronic fatigue syndrome patients

    doi: 10.1186/s12967-021-02974-4

    Figure Lengend Snippet: Co-localisation and immunofluorescent images of TRPM3 with actin in NK cells of HC. A Example of immunostaining of actin (phalloidin, red), nucleus (DAPI, blue), TRPM3 (green) in an NKLa cell under control IL-2 conditions. Cells were stimulated overnight with IL-2 (20 IU) (control) and treated with PregS (30 µM) and ononetin (3 µM). Images taken using Nikon A1R + confocal microscopy. B Bar graphs represent correlation between target antigens using Pearson’s correlation coefficient and the degree of overlap between target antigens using Mander’s overlap coefficient. Co-localisation coefficients K1 (TRPM3) and K2 (actin) were used to determine contribution to co-localisation. Number of cells analysed are included within bar graphs: Control n = 260; PregS n = 277; and PregS + Ono n = 219. Data presented as mean ± SEM and *p

    Article Snippet: Anti-PIP2 was purchased from Abcam (product code: ab11039) while anti-TRPM3 was purchased from Alomone Labs (Product code: ACC-050) and reconstituted at 0.8 mg/ml in distilled water.

    Techniques: Immunostaining, Confocal Microscopy

    Co-localisation and immunofluorescent images of TRPM3 with actin in NK cells of ME/CFS patients. A Example of immunostaining of actin (phalloidin, red), nucleus (DAPI, blue), TRPM3 (green) in NK cell under control IL-2 condition. Cells were stimulated overnight with IL-2 (20 IU) (control) and treated with PregS (30 µM) and ononetin (3 µM). Images taken using Nikon A1R + confocal microscopy. B Bar graphs represent correlation between target antigens using Pearson’s correlation coefficient and the degree of overlap between target antigens using Mander’s overlap coefficient. Co-localisation coefficients K1 (TRPM3) and K2 (actin) were used to determine contribution to co-localisation. Number of cells analysed are included within bar graphs: Control n = 185; PregS n = 193; and PregS + Ono n = 197. Data presented as mean ± SEM. TRPM3, transient receptor potential Melastatin; PregS, pregnenolone sulfate; Ono, ononetin; IL-2, interleukin 2

    Journal: Journal of Translational Medicine

    Article Title: The effect of IL-2 stimulation and treatment of TRPM3 on channel co-localisation with PIP2 and NK cell function in myalgic encephalomyelitis/chronic fatigue syndrome patients

    doi: 10.1186/s12967-021-02974-4

    Figure Lengend Snippet: Co-localisation and immunofluorescent images of TRPM3 with actin in NK cells of ME/CFS patients. A Example of immunostaining of actin (phalloidin, red), nucleus (DAPI, blue), TRPM3 (green) in NK cell under control IL-2 condition. Cells were stimulated overnight with IL-2 (20 IU) (control) and treated with PregS (30 µM) and ononetin (3 µM). Images taken using Nikon A1R + confocal microscopy. B Bar graphs represent correlation between target antigens using Pearson’s correlation coefficient and the degree of overlap between target antigens using Mander’s overlap coefficient. Co-localisation coefficients K1 (TRPM3) and K2 (actin) were used to determine contribution to co-localisation. Number of cells analysed are included within bar graphs: Control n = 185; PregS n = 193; and PregS + Ono n = 197. Data presented as mean ± SEM. TRPM3, transient receptor potential Melastatin; PregS, pregnenolone sulfate; Ono, ononetin; IL-2, interleukin 2

    Article Snippet: Anti-PIP2 was purchased from Abcam (product code: ab11039) while anti-TRPM3 was purchased from Alomone Labs (Product code: ACC-050) and reconstituted at 0.8 mg/ml in distilled water.

    Techniques: Immunostaining, Confocal Microscopy

    Co-localisation and immunofluorescent images of TRPM3 with PIP 2 in NK cells of ME/CFS. patients. A Example of immunostaining of PIP 2 (red), nucleus (DAPI, blue), TRPM3 (green) in NK cell under control IL-2 conditions. Cells were stimulated overnight with IL-2 (20 IU) (control) and treated with PregS (30 µM) and ononetin (3 µM). Images taken using Nikon A1R + confocal microscopy. B Bar graphs represent correlation between target antigens using Pearson’s correlation coefficient and the degree of overlap between target antigens using Mander’s overlap coefficient. Co-localisation coefficients K1 (TRPM3) and K2 (PIP 2 ) were used to determine contribution to co-localisation. Number of cells analysed are included within bar graphs: Control n = 214; PregS n = 225; and PregS + Ono n = 220. Data presented as mean ± SEM and *p

    Journal: Journal of Translational Medicine

    Article Title: The effect of IL-2 stimulation and treatment of TRPM3 on channel co-localisation with PIP2 and NK cell function in myalgic encephalomyelitis/chronic fatigue syndrome patients

    doi: 10.1186/s12967-021-02974-4

    Figure Lengend Snippet: Co-localisation and immunofluorescent images of TRPM3 with PIP 2 in NK cells of ME/CFS. patients. A Example of immunostaining of PIP 2 (red), nucleus (DAPI, blue), TRPM3 (green) in NK cell under control IL-2 conditions. Cells were stimulated overnight with IL-2 (20 IU) (control) and treated with PregS (30 µM) and ononetin (3 µM). Images taken using Nikon A1R + confocal microscopy. B Bar graphs represent correlation between target antigens using Pearson’s correlation coefficient and the degree of overlap between target antigens using Mander’s overlap coefficient. Co-localisation coefficients K1 (TRPM3) and K2 (PIP 2 ) were used to determine contribution to co-localisation. Number of cells analysed are included within bar graphs: Control n = 214; PregS n = 225; and PregS + Ono n = 220. Data presented as mean ± SEM and *p

    Article Snippet: Anti-PIP2 was purchased from Abcam (product code: ab11039) while anti-TRPM3 was purchased from Alomone Labs (Product code: ACC-050) and reconstituted at 0.8 mg/ml in distilled water.

    Techniques: Immunostaining, Confocal Microscopy

    Co-localisation and immunofluorescent images of TRPM3 with PIP 2 in NK cells of HC. A Example of immunostaining of PIP 2 (red), nucleus (DAPI, blue), TRPM3 (green)) in an NK cell under control IL-2 conditions. Images taken using Nikon A1R + confocal microscopy. B Bar graphs represent correlation between target antigens using Pearson’s correlation coefficient and the degree of overlap between target antigens using Mander’s overlap coefficient. Co-localisation coefficients K1 (TRPM3) and K2 (PIP 2 ) were used to determine contribution to co-localisation. Number of cells analysed are included within bar graphs: Control n = 287; PregS n = 292; and PregS + Ono n = 237. Data presented as mean ± SEM and *p

    Journal: Journal of Translational Medicine

    Article Title: The effect of IL-2 stimulation and treatment of TRPM3 on channel co-localisation with PIP2 and NK cell function in myalgic encephalomyelitis/chronic fatigue syndrome patients

    doi: 10.1186/s12967-021-02974-4

    Figure Lengend Snippet: Co-localisation and immunofluorescent images of TRPM3 with PIP 2 in NK cells of HC. A Example of immunostaining of PIP 2 (red), nucleus (DAPI, blue), TRPM3 (green)) in an NK cell under control IL-2 conditions. Images taken using Nikon A1R + confocal microscopy. B Bar graphs represent correlation between target antigens using Pearson’s correlation coefficient and the degree of overlap between target antigens using Mander’s overlap coefficient. Co-localisation coefficients K1 (TRPM3) and K2 (PIP 2 ) were used to determine contribution to co-localisation. Number of cells analysed are included within bar graphs: Control n = 287; PregS n = 292; and PregS + Ono n = 237. Data presented as mean ± SEM and *p

    Article Snippet: Anti-PIP2 was purchased from Abcam (product code: ab11039) while anti-TRPM3 was purchased from Alomone Labs (Product code: ACC-050) and reconstituted at 0.8 mg/ml in distilled water.

    Techniques: Immunostaining, Confocal Microscopy

    Electrophysiological properties of TRPM3 in human stem cell‐derived sensory neurons (hSCDS) neurons. (a) Time course of inward (at −120 mV) and outward (at +80 mV) whole‐cell currents in hSCDS neurons showing the effects of the TRPM3 agonists pregnenolone sulphate (PS; 40 μM) and CIM0216 (1 μM). (b) Quantification of the amplitude of inward and outward currents activated by PS and CIM0216 ( n = ]

    Journal: British Journal of Pharmacology

    Article Title: Functional expression and pharmacological modulation of TRPM3 in human sensory neurons, et al. Functional expression and pharmacological modulation of TRPM3 in human sensory neurons

    doi: 10.1111/bph.14994

    Figure Lengend Snippet: Electrophysiological properties of TRPM3 in human stem cell‐derived sensory neurons (hSCDS) neurons. (a) Time course of inward (at −120 mV) and outward (at +80 mV) whole‐cell currents in hSCDS neurons showing the effects of the TRPM3 agonists pregnenolone sulphate (PS; 40 μM) and CIM0216 (1 μM). (b) Quantification of the amplitude of inward and outward currents activated by PS and CIM0216 ( n = ]

    Article Snippet: After blocking, TRPM3 antibody (Alomone Labs, Cat# ACC‐050, RRID:AB_10918820; 0.2 μg·ml−1 ) was incubated overnight.

    Techniques: Derivative Assay

    Pharmacological characterization of TRPM3 in human stem cell‐derived sensory neurons (hSCDS) neurons. (a) Time course of the intracellular Ca 2+ concentration in PS responding hSCDS neurons upon increasing the concentration of pregnenolone sulphate (PS) from 1 to 300 μM. (b) Concentration dependence of the PS‐induced calcium response in hSCDS neurons. Responses were normalized to the maximal response ( n = 117). Solid line represents a fit using a Hill function. (c) Normalized calcium traces showing the effect of increasing concentrations of the TRPM3 antagonist isosakuranetin (Isoas) on the response to PS (40 μM). Green trace represents the vehicle control, showing mild rundown of the signal in the absence of antagonist. (d) Concentration–response curve for the inhibition of PS‐evoked calcium responses by isosakuranetin ( n = 93; red squares) and primidone ( n = ]

    Journal: British Journal of Pharmacology

    Article Title: Functional expression and pharmacological modulation of TRPM3 in human sensory neurons, et al. Functional expression and pharmacological modulation of TRPM3 in human sensory neurons

    doi: 10.1111/bph.14994

    Figure Lengend Snippet: Pharmacological characterization of TRPM3 in human stem cell‐derived sensory neurons (hSCDS) neurons. (a) Time course of the intracellular Ca 2+ concentration in PS responding hSCDS neurons upon increasing the concentration of pregnenolone sulphate (PS) from 1 to 300 μM. (b) Concentration dependence of the PS‐induced calcium response in hSCDS neurons. Responses were normalized to the maximal response ( n = 117). Solid line represents a fit using a Hill function. (c) Normalized calcium traces showing the effect of increasing concentrations of the TRPM3 antagonist isosakuranetin (Isoas) on the response to PS (40 μM). Green trace represents the vehicle control, showing mild rundown of the signal in the absence of antagonist. (d) Concentration–response curve for the inhibition of PS‐evoked calcium responses by isosakuranetin ( n = 93; red squares) and primidone ( n = ]

    Article Snippet: After blocking, TRPM3 antibody (Alomone Labs, Cat# ACC‐050, RRID:AB_10918820; 0.2 μg·ml−1 ) was incubated overnight.

    Techniques: Derivative Assay, Concentration Assay, Inhibition

    Functional TRPM3 expression in human dorsal root ganglion (DRG) neurons. (a) Expression levels of TRP channel mRNA relative to HPRT in DRG neurons from two donors, determined using RT‐qPCR. Access to fresh human DRG tissue is scarce, hence the limited number of samples in this exploratory experiment. nd, not detected. (b) Representative example of changes in Fluo‐8‐fluorescence in human DRG neurons in response to the TRPM3 agonists pregnenolone sulphate (PS; 50 μM) and CIM0216 (10 μM) and to the TRPV1 agonist capsaicin (Caps; 200 nM). A high K + . (c) Pie chart showing the distribution of neurons responding to PS, capsaicin or both ( n . (e) Normalized responses to repeated PS applications. Neurons were stimulated three times with PS, and the second application occurred in the presence of either isosakuranetin (10 μM; n = 50) or vehicle control ( n = 39). * P ]

    Journal: British Journal of Pharmacology

    Article Title: Functional expression and pharmacological modulation of TRPM3 in human sensory neurons, et al. Functional expression and pharmacological modulation of TRPM3 in human sensory neurons

    doi: 10.1111/bph.14994

    Figure Lengend Snippet: Functional TRPM3 expression in human dorsal root ganglion (DRG) neurons. (a) Expression levels of TRP channel mRNA relative to HPRT in DRG neurons from two donors, determined using RT‐qPCR. Access to fresh human DRG tissue is scarce, hence the limited number of samples in this exploratory experiment. nd, not detected. (b) Representative example of changes in Fluo‐8‐fluorescence in human DRG neurons in response to the TRPM3 agonists pregnenolone sulphate (PS; 50 μM) and CIM0216 (10 μM) and to the TRPV1 agonist capsaicin (Caps; 200 nM). A high K + . (c) Pie chart showing the distribution of neurons responding to PS, capsaicin or both ( n . (e) Normalized responses to repeated PS applications. Neurons were stimulated three times with PS, and the second application occurred in the presence of either isosakuranetin (10 μM; n = 50) or vehicle control ( n = 39). * P ]

    Article Snippet: After blocking, TRPM3 antibody (Alomone Labs, Cat# ACC‐050, RRID:AB_10918820; 0.2 μg·ml−1 ) was incubated overnight.

    Techniques: Functional Assay, Expressing, Quantitative RT-PCR, Fluorescence

    Modulation of TRPM3 in human stem cell‐derived sensory neurons (hSCDS) neurons by GPCRs. (a) Example intracellular calcium measurements showing variable degrees of inhibition of PS‐induced responses upon activation of endogenously expressed μ‐opioid receptors using DAMGO (300 nM). (b) Scatter plot showing the range of DAMGO‐induced inhibition of PS responses in hSCDS neurons. Red line represents the mean percentage of TRPM3 inhibition caused by DAMGO (data from six experiments, with hSCDS neurons from three different differentiations). (c) Example intracellular calcium traces showing inhibition of PS‐induced responses by the GABA B ]

    Journal: British Journal of Pharmacology

    Article Title: Functional expression and pharmacological modulation of TRPM3 in human sensory neurons, et al. Functional expression and pharmacological modulation of TRPM3 in human sensory neurons

    doi: 10.1111/bph.14994

    Figure Lengend Snippet: Modulation of TRPM3 in human stem cell‐derived sensory neurons (hSCDS) neurons by GPCRs. (a) Example intracellular calcium measurements showing variable degrees of inhibition of PS‐induced responses upon activation of endogenously expressed μ‐opioid receptors using DAMGO (300 nM). (b) Scatter plot showing the range of DAMGO‐induced inhibition of PS responses in hSCDS neurons. Red line represents the mean percentage of TRPM3 inhibition caused by DAMGO (data from six experiments, with hSCDS neurons from three different differentiations). (c) Example intracellular calcium traces showing inhibition of PS‐induced responses by the GABA B ]

    Article Snippet: After blocking, TRPM3 antibody (Alomone Labs, Cat# ACC‐050, RRID:AB_10918820; 0.2 μg·ml−1 ) was incubated overnight.

    Techniques: Derivative Assay, Inhibition, Activation Assay

    Functional expression profile of somatosensory TRP channels in human stem cell‐derived sensory neurons (hSCDS) neurons. (a) Changes in intracellular calcium in single hSCDS neurons stimulated with the TRP channel agonists menthol (100 μM), MO (100 μM), capsaicin (Caps;1 μM), and pregnenolone sulphate (PS;40 μM) and with a high K + solution to probe excitability. (b) Venn diagram showing the pattern of responses to TRP channel agonists in hSCDS neurons ( n = 1,180 cells in five independent differentiations). (c) Intracellular calcium measurements showing reversible inhibition of PS‐evoked responses by isosakuranetin (5 μM). PS‐responsive hSCDS neurons also responded to the synthetic TRPM3 agonist CIM0216 (1 μM). (d) Quantification of calcium responses for experiments as in panel (c) ( n = ]

    Journal: British Journal of Pharmacology

    Article Title: Functional expression and pharmacological modulation of TRPM3 in human sensory neurons, et al. Functional expression and pharmacological modulation of TRPM3 in human sensory neurons

    doi: 10.1111/bph.14994

    Figure Lengend Snippet: Functional expression profile of somatosensory TRP channels in human stem cell‐derived sensory neurons (hSCDS) neurons. (a) Changes in intracellular calcium in single hSCDS neurons stimulated with the TRP channel agonists menthol (100 μM), MO (100 μM), capsaicin (Caps;1 μM), and pregnenolone sulphate (PS;40 μM) and with a high K + solution to probe excitability. (b) Venn diagram showing the pattern of responses to TRP channel agonists in hSCDS neurons ( n = 1,180 cells in five independent differentiations). (c) Intracellular calcium measurements showing reversible inhibition of PS‐evoked responses by isosakuranetin (5 μM). PS‐responsive hSCDS neurons also responded to the synthetic TRPM3 agonist CIM0216 (1 μM). (d) Quantification of calcium responses for experiments as in panel (c) ( n = ]

    Article Snippet: After blocking, TRPM3 antibody (Alomone Labs, Cat# ACC‐050, RRID:AB_10918820; 0.2 μg·ml−1 ) was incubated overnight.

    Techniques: Functional Assay, Expressing, Derivative Assay, Inhibition