anti v1ar  (Alomone Labs)


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    Alomone Labs anti v1ar
    Anti V1ar, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 91 stars, based on 1 article reviews
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    anti v1ar  (Alomone Labs)


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    Alomone Labs anti v1ar
    Anti V1ar, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti v1ar/product/Alomone Labs
    Average 91 stars, based on 1 article reviews
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    anti v1ar antibody  (Alomone Labs)


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  • 91

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    Alomone Labs anti v1ar antibody
    Anti V1ar Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti v1ar antibody/product/Alomone Labs
    Average 91 stars, based on 1 article reviews
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    anti v1ar antibody - by Bioz Stars, 2023-01
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    anti v1ar  (Alomone Labs)


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    Alomone Labs anti v1ar
    Anti V1ar, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti v1ar/product/Alomone Labs
    Average 91 stars, based on 1 article reviews
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    v1ar  (Alomone Labs)


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    Alomone Labs v1ar
    V1ar, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/v1ar/product/Alomone Labs
    Average 91 stars, based on 1 article reviews
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    v1ar - by Bioz Stars, 2023-01
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    rabbit anti v1a receptor polyclonal antibody  (Alomone Labs)


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    Alomone Labs rabbit anti v1a receptor polyclonal antibody
    (A) Thermal paw-withdrawal latency was measured and compared before the injection with 30 min and 60 min after the IP injection of vehicle, OT (1 mg/kg) and OT with OTRA (A selective oxytocin receptor antagonist 1 mg/kg), atosiban (an oxytocin and vasopressin receptor antagonist, 1 mg/kg) and V1aRA (a selective <t>vasopressin</t> <t>1a</t> receptor antagonist, 1 mg/kg) in Hargreaves' test. ** p<0.001, *** p<0.0001, Two-way RM ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM (n=6, each group). (B) Nocifensive responses at 60 min after IP co-injection of antagonists were compared to the OT injection. ** p<0.001, *** p<0.0001, One-way ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. (n=6, each group). OT, oxytocin; V1aRA, vasopressin-1a receptor antagonist; OTRA, oxytocin receptor antagonist; ns, not significant.
    Rabbit Anti V1a Receptor Polyclonal Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti v1a receptor polyclonal antibody/product/Alomone Labs
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    91/100 stars

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    1) Product Images from "Oxytocin produces thermal analgesia via vasopressin-1a receptor by modulating TRPV1 and potassium conductance in the dorsal root ganglion neurons"

    Article Title: Oxytocin produces thermal analgesia via vasopressin-1a receptor by modulating TRPV1 and potassium conductance in the dorsal root ganglion neurons

    Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

    doi: 10.4196/kjpp.2018.22.2.173

    (A) Thermal paw-withdrawal latency was measured and compared before the injection with 30 min and 60 min after the IP injection of vehicle, OT (1 mg/kg) and OT with OTRA (A selective oxytocin receptor antagonist 1 mg/kg), atosiban (an oxytocin and vasopressin receptor antagonist, 1 mg/kg) and V1aRA (a selective vasopressin 1a receptor antagonist, 1 mg/kg) in Hargreaves' test. ** p<0.001, *** p<0.0001, Two-way RM ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM (n=6, each group). (B) Nocifensive responses at 60 min after IP co-injection of antagonists were compared to the OT injection. ** p<0.001, *** p<0.0001, One-way ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. (n=6, each group). OT, oxytocin; V1aRA, vasopressin-1a receptor antagonist; OTRA, oxytocin receptor antagonist; ns, not significant.
    Figure Legend Snippet: (A) Thermal paw-withdrawal latency was measured and compared before the injection with 30 min and 60 min after the IP injection of vehicle, OT (1 mg/kg) and OT with OTRA (A selective oxytocin receptor antagonist 1 mg/kg), atosiban (an oxytocin and vasopressin receptor antagonist, 1 mg/kg) and V1aRA (a selective vasopressin 1a receptor antagonist, 1 mg/kg) in Hargreaves' test. ** p<0.001, *** p<0.0001, Two-way RM ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM (n=6, each group). (B) Nocifensive responses at 60 min after IP co-injection of antagonists were compared to the OT injection. ** p<0.001, *** p<0.0001, One-way ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. (n=6, each group). OT, oxytocin; V1aRA, vasopressin-1a receptor antagonist; OTRA, oxytocin receptor antagonist; ns, not significant.

    Techniques Used: Injection

    Representative traces of intracellular calcium responses of cultured sensory neurons to 3 repetitive application of (A) 200 nM CAP for 10 s, (B) repetitive CAP application with pre-application of 5 µM of oxytocin for 120 s, (C) CAP application with 5 µM of oxytocin mixed with 20 µM of atosiban, and (D) 5 µM of oxytocin mixed with 5 µM of V1aRA before the second capsaicin application. (E) The proportion of oxytocin responsive cells within the DRG neurons responding to capsaicin in calcium imaging. (F) Summary of normalized ratio of capsaicin responses by without OT (n=21), OT alone (n=70), and OT with its antagonists, ATO (n=66) and V1aRA (n=70), relative to peak amplitude of 1st CAP transient. Before CAP application, 10 µM PMA was mixed with CAP solution. * p<0.05, One-way ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. CAP, capsaicin; PMA, Phorbol 12-myristate 13-acetate; OT, oxytocin; ATO, atosiban; V1aRA, vasopressin-1a receptor antagonist; ns, not significant.
    Figure Legend Snippet: Representative traces of intracellular calcium responses of cultured sensory neurons to 3 repetitive application of (A) 200 nM CAP for 10 s, (B) repetitive CAP application with pre-application of 5 µM of oxytocin for 120 s, (C) CAP application with 5 µM of oxytocin mixed with 20 µM of atosiban, and (D) 5 µM of oxytocin mixed with 5 µM of V1aRA before the second capsaicin application. (E) The proportion of oxytocin responsive cells within the DRG neurons responding to capsaicin in calcium imaging. (F) Summary of normalized ratio of capsaicin responses by without OT (n=21), OT alone (n=70), and OT with its antagonists, ATO (n=66) and V1aRA (n=70), relative to peak amplitude of 1st CAP transient. Before CAP application, 10 µM PMA was mixed with CAP solution. * p<0.05, One-way ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. CAP, capsaicin; PMA, Phorbol 12-myristate 13-acetate; OT, oxytocin; ATO, atosiban; V1aRA, vasopressin-1a receptor antagonist; ns, not significant.

    Techniques Used: Cell Culture, Imaging

    (A, left panel) Representative recording illustrating effect of 5 µM oxytocin on action potential firing. The effect of oxytocin was reversed after 5-min washout. (A, right panel) Oxytocin decreased the mean number of action potential evoked by current injection (n=12). (B) 20 µM of Atosiban (n=9) and (C) 5 µM of V1aRA (n=9) reversed oxytocin-induced reduction of the number of action potentials in small to medium-sized DRG neurons. * p<0.01, *** p<0.0001, repeated measured-ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. OT, Oxytocin; V1aRA, vasopressin-1a receptor antagonist; ns, not significant.
    Figure Legend Snippet: (A, left panel) Representative recording illustrating effect of 5 µM oxytocin on action potential firing. The effect of oxytocin was reversed after 5-min washout. (A, right panel) Oxytocin decreased the mean number of action potential evoked by current injection (n=12). (B) 20 µM of Atosiban (n=9) and (C) 5 µM of V1aRA (n=9) reversed oxytocin-induced reduction of the number of action potentials in small to medium-sized DRG neurons. * p<0.01, *** p<0.0001, repeated measured-ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. OT, Oxytocin; V1aRA, vasopressin-1a receptor antagonist; ns, not significant.

    Techniques Used: Injection

    (A) Depolarizing (from –110 mV to 0 mV) ramp protocol for voltage-clamp experiments (B, left panel) Representative trace showing effects of oxytocin (5 µM) on I-V relationship response to ramp depolarization. (B, middle panel) Representative trace illustrating markedly enhanced a voltage-activated outward current and slightly enhanced a hyperpolarization-activated inward current (B, right panel) Oxytocin increased the mean peak current density at 0 mV by inducing outward potassium current in small to medium-sized DRG neurons (n=20). (C) 20 µM of atosiban (n=24) and (D) 5 µM of V1aRA blocked oxytocin-induced the voltage-activated outward potassium current and the hyperpolarization-activated inward current. TTX (0.5 µM) were treated in solution of all experiments (n=13). * p<0.01, ** p<0.001, repeated measured-ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. OT, Oxytocin; V1aRA, vasopressin-1a receptor antagonist; ns, not significant.
    Figure Legend Snippet: (A) Depolarizing (from –110 mV to 0 mV) ramp protocol for voltage-clamp experiments (B, left panel) Representative trace showing effects of oxytocin (5 µM) on I-V relationship response to ramp depolarization. (B, middle panel) Representative trace illustrating markedly enhanced a voltage-activated outward current and slightly enhanced a hyperpolarization-activated inward current (B, right panel) Oxytocin increased the mean peak current density at 0 mV by inducing outward potassium current in small to medium-sized DRG neurons (n=20). (C) 20 µM of atosiban (n=24) and (D) 5 µM of V1aRA blocked oxytocin-induced the voltage-activated outward potassium current and the hyperpolarization-activated inward current. TTX (0.5 µM) were treated in solution of all experiments (n=13). * p<0.01, ** p<0.001, repeated measured-ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. OT, Oxytocin; V1aRA, vasopressin-1a receptor antagonist; ns, not significant.

    Techniques Used:

    vasopressin receptor 1a  (Alomone Labs)


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    Alomone Labs vasopressin receptor 1a
    a Serial coronal sections showing the ERα immunolabelling at the Bregma rostro-caudal coordinates (numbers in mm under the photomicrographs). Boxed area in A at higher magnification showing the exclusive nuclear labeling pattern. The bold numbered levels (A 1 and A 8 ), were chosen to show that no positive labeling was found in either rostral or caudal directions. A 9 : a horizontal view of the distribution of estrogen-receptive cells was symbolized by the red oval. A 10 : sagittal view of rat brain atlas, modified from Paxinos & Watson , at lat. 0.90 mm, where lateral habenula (LHb) is symbolized with a gray shade and A9 plane was symbolized with a horizontal line. b In situ hybridization using multiple RNAscope methods. B 1 : multiplex fluorescence method, Esr1, gene that encodes ERα (red punctuated labeling) co-expressed with Slc32a1, gene that encodes VGAT (green punctuated labeling); arrows indicate the double-labeled cells; B 2 : duplex method, Esr1 (red punctuated labeling) co-localization with Slc17a6, gene that encodes VGLUT2 (green punctuated labeling); arrows indicate the double-labeled cells; B 3 : with duplex method, Slc32a1 encoding VGAT (red punctuated labeling) shows complete co-localization with Slc17a6 encoding VGLUT2 (green punctuated labeling); arrows indicate the double-labeled cells. Inset of B 3 , Slc32a1 expression in a sexually active (SA) rat LHb, Br. −3.72 mm (brown labeling, single chromogenic-Brown method-RNAscope). *Note the similarity with ERα expression in A . c Indirect immunohistochemistry showing the GABAergic nature of the ERα+ neurons (red) in a SA rat brain. The GABA antibody (green, Sigma, A0310) produced characteristic surface labeling (C 1 , C 2 : the strip-like image was produced by Vibratome slicing leaving the brain section with an uneven surface). d ERα+ neurons co-express receptor/receptor subtypes for vasopressin, orexin, dopamine, and serotonin. D 1 : In situ hybridization using RNAscope-multiplex method targeting Esr1 (red dots), Slc32a1 (white dots) and Hcrtr2, gene encodes the orexin receptor 2 (green dots). D 2 , D 3 : Indirect immunofluorescence reactions, showing that the ERα-IR cells co-expressed vasopressin receptor <t>V1a</t> (inset showing the V1a antibody labeling pattern in temporal hippocampus CA2 cells body layer. See also SI Fig. for more information about this antibody), dopamine receptor D5R (also called D1Rb) and serotonin receptor 5-HTr2a, respectively. Scale bars: A 5 and b : 500 µm and rest: 10 µm
    Vasopressin Receptor 1a, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "A GABAergic cell type in the lateral habenula links hypothalamic homeostatic and midbrain motivation circuits with sex steroid signaling"

    Article Title: A GABAergic cell type in the lateral habenula links hypothalamic homeostatic and midbrain motivation circuits with sex steroid signaling

    Journal: Translational Psychiatry

    doi: 10.1038/s41398-018-0099-5

    a Serial coronal sections showing the ERα immunolabelling at the Bregma rostro-caudal coordinates (numbers in mm under the photomicrographs). Boxed area in A at higher magnification showing the exclusive nuclear labeling pattern. The bold numbered levels (A 1 and A 8 ), were chosen to show that no positive labeling was found in either rostral or caudal directions. A 9 : a horizontal view of the distribution of estrogen-receptive cells was symbolized by the red oval. A 10 : sagittal view of rat brain atlas, modified from Paxinos & Watson , at lat. 0.90 mm, where lateral habenula (LHb) is symbolized with a gray shade and A9 plane was symbolized with a horizontal line. b In situ hybridization using multiple RNAscope methods. B 1 : multiplex fluorescence method, Esr1, gene that encodes ERα (red punctuated labeling) co-expressed with Slc32a1, gene that encodes VGAT (green punctuated labeling); arrows indicate the double-labeled cells; B 2 : duplex method, Esr1 (red punctuated labeling) co-localization with Slc17a6, gene that encodes VGLUT2 (green punctuated labeling); arrows indicate the double-labeled cells; B 3 : with duplex method, Slc32a1 encoding VGAT (red punctuated labeling) shows complete co-localization with Slc17a6 encoding VGLUT2 (green punctuated labeling); arrows indicate the double-labeled cells. Inset of B 3 , Slc32a1 expression in a sexually active (SA) rat LHb, Br. −3.72 mm (brown labeling, single chromogenic-Brown method-RNAscope). *Note the similarity with ERα expression in A . c Indirect immunohistochemistry showing the GABAergic nature of the ERα+ neurons (red) in a SA rat brain. The GABA antibody (green, Sigma, A0310) produced characteristic surface labeling (C 1 , C 2 : the strip-like image was produced by Vibratome slicing leaving the brain section with an uneven surface). d ERα+ neurons co-express receptor/receptor subtypes for vasopressin, orexin, dopamine, and serotonin. D 1 : In situ hybridization using RNAscope-multiplex method targeting Esr1 (red dots), Slc32a1 (white dots) and Hcrtr2, gene encodes the orexin receptor 2 (green dots). D 2 , D 3 : Indirect immunofluorescence reactions, showing that the ERα-IR cells co-expressed vasopressin receptor V1a (inset showing the V1a antibody labeling pattern in temporal hippocampus CA2 cells body layer. See also SI Fig. for more information about this antibody), dopamine receptor D5R (also called D1Rb) and serotonin receptor 5-HTr2a, respectively. Scale bars: A 5 and b : 500 µm and rest: 10 µm
    Figure Legend Snippet: a Serial coronal sections showing the ERα immunolabelling at the Bregma rostro-caudal coordinates (numbers in mm under the photomicrographs). Boxed area in A at higher magnification showing the exclusive nuclear labeling pattern. The bold numbered levels (A 1 and A 8 ), were chosen to show that no positive labeling was found in either rostral or caudal directions. A 9 : a horizontal view of the distribution of estrogen-receptive cells was symbolized by the red oval. A 10 : sagittal view of rat brain atlas, modified from Paxinos & Watson , at lat. 0.90 mm, where lateral habenula (LHb) is symbolized with a gray shade and A9 plane was symbolized with a horizontal line. b In situ hybridization using multiple RNAscope methods. B 1 : multiplex fluorescence method, Esr1, gene that encodes ERα (red punctuated labeling) co-expressed with Slc32a1, gene that encodes VGAT (green punctuated labeling); arrows indicate the double-labeled cells; B 2 : duplex method, Esr1 (red punctuated labeling) co-localization with Slc17a6, gene that encodes VGLUT2 (green punctuated labeling); arrows indicate the double-labeled cells; B 3 : with duplex method, Slc32a1 encoding VGAT (red punctuated labeling) shows complete co-localization with Slc17a6 encoding VGLUT2 (green punctuated labeling); arrows indicate the double-labeled cells. Inset of B 3 , Slc32a1 expression in a sexually active (SA) rat LHb, Br. −3.72 mm (brown labeling, single chromogenic-Brown method-RNAscope). *Note the similarity with ERα expression in A . c Indirect immunohistochemistry showing the GABAergic nature of the ERα+ neurons (red) in a SA rat brain. The GABA antibody (green, Sigma, A0310) produced characteristic surface labeling (C 1 , C 2 : the strip-like image was produced by Vibratome slicing leaving the brain section with an uneven surface). d ERα+ neurons co-express receptor/receptor subtypes for vasopressin, orexin, dopamine, and serotonin. D 1 : In situ hybridization using RNAscope-multiplex method targeting Esr1 (red dots), Slc32a1 (white dots) and Hcrtr2, gene encodes the orexin receptor 2 (green dots). D 2 , D 3 : Indirect immunofluorescence reactions, showing that the ERα-IR cells co-expressed vasopressin receptor V1a (inset showing the V1a antibody labeling pattern in temporal hippocampus CA2 cells body layer. See also SI Fig. for more information about this antibody), dopamine receptor D5R (also called D1Rb) and serotonin receptor 5-HTr2a, respectively. Scale bars: A 5 and b : 500 µm and rest: 10 µm

    Techniques Used: Labeling, Modification, In Situ Hybridization, Multiplex Assay, Fluorescence, Expressing, Immunohistochemistry, Produced, Stripping Membranes, Immunofluorescence, Antibody Labeling

    avr  (Alomone Labs)


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    Alomone Labs avr
    Avr, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs anti v1ar
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    (A) Thermal paw-withdrawal latency was measured and compared before the injection with 30 min and 60 min after the IP injection of vehicle, OT (1 mg/kg) and OT with OTRA (A selective oxytocin receptor antagonist 1 mg/kg), atosiban (an oxytocin and vasopressin receptor antagonist, 1 mg/kg) and V1aRA (a selective <t>vasopressin</t> <t>1a</t> receptor antagonist, 1 mg/kg) in Hargreaves' test. ** p<0.001, *** p<0.0001, Two-way RM ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM (n=6, each group). (B) Nocifensive responses at 60 min after IP co-injection of antagonists were compared to the OT injection. ** p<0.001, *** p<0.0001, One-way ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. (n=6, each group). OT, oxytocin; V1aRA, vasopressin-1a receptor antagonist; OTRA, oxytocin receptor antagonist; ns, not significant.
    Rabbit Anti V1a Receptor Polyclonal Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti v1a receptor polyclonal antibody/product/Alomone Labs
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    Alomone Labs vasopressin receptor 1a
    a Serial coronal sections showing the ERα immunolabelling at the Bregma rostro-caudal coordinates (numbers in mm under the photomicrographs). Boxed area in A at higher magnification showing the exclusive nuclear labeling pattern. The bold numbered levels (A 1 and A 8 ), were chosen to show that no positive labeling was found in either rostral or caudal directions. A 9 : a horizontal view of the distribution of estrogen-receptive cells was symbolized by the red oval. A 10 : sagittal view of rat brain atlas, modified from Paxinos & Watson , at lat. 0.90 mm, where lateral habenula (LHb) is symbolized with a gray shade and A9 plane was symbolized with a horizontal line. b In situ hybridization using multiple RNAscope methods. B 1 : multiplex fluorescence method, Esr1, gene that encodes ERα (red punctuated labeling) co-expressed with Slc32a1, gene that encodes VGAT (green punctuated labeling); arrows indicate the double-labeled cells; B 2 : duplex method, Esr1 (red punctuated labeling) co-localization with Slc17a6, gene that encodes VGLUT2 (green punctuated labeling); arrows indicate the double-labeled cells; B 3 : with duplex method, Slc32a1 encoding VGAT (red punctuated labeling) shows complete co-localization with Slc17a6 encoding VGLUT2 (green punctuated labeling); arrows indicate the double-labeled cells. Inset of B 3 , Slc32a1 expression in a sexually active (SA) rat LHb, Br. −3.72 mm (brown labeling, single chromogenic-Brown method-RNAscope). *Note the similarity with ERα expression in A . c Indirect immunohistochemistry showing the GABAergic nature of the ERα+ neurons (red) in a SA rat brain. The GABA antibody (green, Sigma, A0310) produced characteristic surface labeling (C 1 , C 2 : the strip-like image was produced by Vibratome slicing leaving the brain section with an uneven surface). d ERα+ neurons co-express receptor/receptor subtypes for vasopressin, orexin, dopamine, and serotonin. D 1 : In situ hybridization using RNAscope-multiplex method targeting Esr1 (red dots), Slc32a1 (white dots) and Hcrtr2, gene encodes the orexin receptor 2 (green dots). D 2 , D 3 : Indirect immunofluorescence reactions, showing that the ERα-IR cells co-expressed vasopressin receptor <t>V1a</t> (inset showing the V1a antibody labeling pattern in temporal hippocampus CA2 cells body layer. See also SI Fig. for more information about this antibody), dopamine receptor D5R (also called D1Rb) and serotonin receptor 5-HTr2a, respectively. Scale bars: A 5 and b : 500 µm and rest: 10 µm
    Vasopressin Receptor 1a, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    a Serial coronal sections showing the ERα immunolabelling at the Bregma rostro-caudal coordinates (numbers in mm under the photomicrographs). Boxed area in A at higher magnification showing the exclusive nuclear labeling pattern. The bold numbered levels (A 1 and A 8 ), were chosen to show that no positive labeling was found in either rostral or caudal directions. A 9 : a horizontal view of the distribution of estrogen-receptive cells was symbolized by the red oval. A 10 : sagittal view of rat brain atlas, modified from Paxinos & Watson , at lat. 0.90 mm, where lateral habenula (LHb) is symbolized with a gray shade and A9 plane was symbolized with a horizontal line. b In situ hybridization using multiple RNAscope methods. B 1 : multiplex fluorescence method, Esr1, gene that encodes ERα (red punctuated labeling) co-expressed with Slc32a1, gene that encodes VGAT (green punctuated labeling); arrows indicate the double-labeled cells; B 2 : duplex method, Esr1 (red punctuated labeling) co-localization with Slc17a6, gene that encodes VGLUT2 (green punctuated labeling); arrows indicate the double-labeled cells; B 3 : with duplex method, Slc32a1 encoding VGAT (red punctuated labeling) shows complete co-localization with Slc17a6 encoding VGLUT2 (green punctuated labeling); arrows indicate the double-labeled cells. Inset of B 3 , Slc32a1 expression in a sexually active (SA) rat LHb, Br. −3.72 mm (brown labeling, single chromogenic-Brown method-RNAscope). *Note the similarity with ERα expression in A . c Indirect immunohistochemistry showing the GABAergic nature of the ERα+ neurons (red) in a SA rat brain. The GABA antibody (green, Sigma, A0310) produced characteristic surface labeling (C 1 , C 2 : the strip-like image was produced by Vibratome slicing leaving the brain section with an uneven surface). d ERα+ neurons co-express receptor/receptor subtypes for vasopressin, orexin, dopamine, and serotonin. D 1 : In situ hybridization using RNAscope-multiplex method targeting Esr1 (red dots), Slc32a1 (white dots) and Hcrtr2, gene encodes the orexin receptor 2 (green dots). D 2 , D 3 : Indirect immunofluorescence reactions, showing that the ERα-IR cells co-expressed vasopressin receptor <t>V1a</t> (inset showing the V1a antibody labeling pattern in temporal hippocampus CA2 cells body layer. See also SI Fig. for more information about this antibody), dopamine receptor D5R (also called D1Rb) and serotonin receptor 5-HTr2a, respectively. Scale bars: A 5 and b : 500 µm and rest: 10 µm
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    (A) Thermal paw-withdrawal latency was measured and compared before the injection with 30 min and 60 min after the IP injection of vehicle, OT (1 mg/kg) and OT with OTRA (A selective oxytocin receptor antagonist 1 mg/kg), atosiban (an oxytocin and vasopressin receptor antagonist, 1 mg/kg) and V1aRA (a selective vasopressin 1a receptor antagonist, 1 mg/kg) in Hargreaves' test. ** p<0.001, *** p<0.0001, Two-way RM ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM (n=6, each group). (B) Nocifensive responses at 60 min after IP co-injection of antagonists were compared to the OT injection. ** p<0.001, *** p<0.0001, One-way ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. (n=6, each group). OT, oxytocin; V1aRA, vasopressin-1a receptor antagonist; OTRA, oxytocin receptor antagonist; ns, not significant.

    Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

    Article Title: Oxytocin produces thermal analgesia via vasopressin-1a receptor by modulating TRPV1 and potassium conductance in the dorsal root ganglion neurons

    doi: 10.4196/kjpp.2018.22.2.173

    Figure Lengend Snippet: (A) Thermal paw-withdrawal latency was measured and compared before the injection with 30 min and 60 min after the IP injection of vehicle, OT (1 mg/kg) and OT with OTRA (A selective oxytocin receptor antagonist 1 mg/kg), atosiban (an oxytocin and vasopressin receptor antagonist, 1 mg/kg) and V1aRA (a selective vasopressin 1a receptor antagonist, 1 mg/kg) in Hargreaves' test. ** p<0.001, *** p<0.0001, Two-way RM ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM (n=6, each group). (B) Nocifensive responses at 60 min after IP co-injection of antagonists were compared to the OT injection. ** p<0.001, *** p<0.0001, One-way ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. (n=6, each group). OT, oxytocin; V1aRA, vasopressin-1a receptor antagonist; OTRA, oxytocin receptor antagonist; ns, not significant.

    Article Snippet: The primary antibodies were rabbit anti-V1a receptor polyclonal antibody (1:400; Alomone Labs, UK) and guinea pig anti-TRPV1 polyclonal antibody (1:500; Novus biologicals, USA), diluted in PBS containing 0.3% Triton X-100, 1% normal donkey serum (Jackson ImmunoResearch Co, USA), incubated overnight at 4℃ then incubated for 1 h with donkey anti-guinea pig IgG conjugated with cyanine dye 3 (Cy3, 1:200; Jackson ImmunoResearch Co, USA) and donkey anti-rabbit IgG conjugated with fluorescein-isothiocyanate (FITC, 1:200; Jackson ImmunoResearch Co, USA) diluted in the same buffer as the primary antibody.

    Techniques: Injection

    Representative traces of intracellular calcium responses of cultured sensory neurons to 3 repetitive application of (A) 200 nM CAP for 10 s, (B) repetitive CAP application with pre-application of 5 µM of oxytocin for 120 s, (C) CAP application with 5 µM of oxytocin mixed with 20 µM of atosiban, and (D) 5 µM of oxytocin mixed with 5 µM of V1aRA before the second capsaicin application. (E) The proportion of oxytocin responsive cells within the DRG neurons responding to capsaicin in calcium imaging. (F) Summary of normalized ratio of capsaicin responses by without OT (n=21), OT alone (n=70), and OT with its antagonists, ATO (n=66) and V1aRA (n=70), relative to peak amplitude of 1st CAP transient. Before CAP application, 10 µM PMA was mixed with CAP solution. * p<0.05, One-way ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. CAP, capsaicin; PMA, Phorbol 12-myristate 13-acetate; OT, oxytocin; ATO, atosiban; V1aRA, vasopressin-1a receptor antagonist; ns, not significant.

    Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

    Article Title: Oxytocin produces thermal analgesia via vasopressin-1a receptor by modulating TRPV1 and potassium conductance in the dorsal root ganglion neurons

    doi: 10.4196/kjpp.2018.22.2.173

    Figure Lengend Snippet: Representative traces of intracellular calcium responses of cultured sensory neurons to 3 repetitive application of (A) 200 nM CAP for 10 s, (B) repetitive CAP application with pre-application of 5 µM of oxytocin for 120 s, (C) CAP application with 5 µM of oxytocin mixed with 20 µM of atosiban, and (D) 5 µM of oxytocin mixed with 5 µM of V1aRA before the second capsaicin application. (E) The proportion of oxytocin responsive cells within the DRG neurons responding to capsaicin in calcium imaging. (F) Summary of normalized ratio of capsaicin responses by without OT (n=21), OT alone (n=70), and OT with its antagonists, ATO (n=66) and V1aRA (n=70), relative to peak amplitude of 1st CAP transient. Before CAP application, 10 µM PMA was mixed with CAP solution. * p<0.05, One-way ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. CAP, capsaicin; PMA, Phorbol 12-myristate 13-acetate; OT, oxytocin; ATO, atosiban; V1aRA, vasopressin-1a receptor antagonist; ns, not significant.

    Article Snippet: The primary antibodies were rabbit anti-V1a receptor polyclonal antibody (1:400; Alomone Labs, UK) and guinea pig anti-TRPV1 polyclonal antibody (1:500; Novus biologicals, USA), diluted in PBS containing 0.3% Triton X-100, 1% normal donkey serum (Jackson ImmunoResearch Co, USA), incubated overnight at 4℃ then incubated for 1 h with donkey anti-guinea pig IgG conjugated with cyanine dye 3 (Cy3, 1:200; Jackson ImmunoResearch Co, USA) and donkey anti-rabbit IgG conjugated with fluorescein-isothiocyanate (FITC, 1:200; Jackson ImmunoResearch Co, USA) diluted in the same buffer as the primary antibody.

    Techniques: Cell Culture, Imaging

    (A, left panel) Representative recording illustrating effect of 5 µM oxytocin on action potential firing. The effect of oxytocin was reversed after 5-min washout. (A, right panel) Oxytocin decreased the mean number of action potential evoked by current injection (n=12). (B) 20 µM of Atosiban (n=9) and (C) 5 µM of V1aRA (n=9) reversed oxytocin-induced reduction of the number of action potentials in small to medium-sized DRG neurons. * p<0.01, *** p<0.0001, repeated measured-ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. OT, Oxytocin; V1aRA, vasopressin-1a receptor antagonist; ns, not significant.

    Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

    Article Title: Oxytocin produces thermal analgesia via vasopressin-1a receptor by modulating TRPV1 and potassium conductance in the dorsal root ganglion neurons

    doi: 10.4196/kjpp.2018.22.2.173

    Figure Lengend Snippet: (A, left panel) Representative recording illustrating effect of 5 µM oxytocin on action potential firing. The effect of oxytocin was reversed after 5-min washout. (A, right panel) Oxytocin decreased the mean number of action potential evoked by current injection (n=12). (B) 20 µM of Atosiban (n=9) and (C) 5 µM of V1aRA (n=9) reversed oxytocin-induced reduction of the number of action potentials in small to medium-sized DRG neurons. * p<0.01, *** p<0.0001, repeated measured-ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. OT, Oxytocin; V1aRA, vasopressin-1a receptor antagonist; ns, not significant.

    Article Snippet: The primary antibodies were rabbit anti-V1a receptor polyclonal antibody (1:400; Alomone Labs, UK) and guinea pig anti-TRPV1 polyclonal antibody (1:500; Novus biologicals, USA), diluted in PBS containing 0.3% Triton X-100, 1% normal donkey serum (Jackson ImmunoResearch Co, USA), incubated overnight at 4℃ then incubated for 1 h with donkey anti-guinea pig IgG conjugated with cyanine dye 3 (Cy3, 1:200; Jackson ImmunoResearch Co, USA) and donkey anti-rabbit IgG conjugated with fluorescein-isothiocyanate (FITC, 1:200; Jackson ImmunoResearch Co, USA) diluted in the same buffer as the primary antibody.

    Techniques: Injection

    (A) Depolarizing (from –110 mV to 0 mV) ramp protocol for voltage-clamp experiments (B, left panel) Representative trace showing effects of oxytocin (5 µM) on I-V relationship response to ramp depolarization. (B, middle panel) Representative trace illustrating markedly enhanced a voltage-activated outward current and slightly enhanced a hyperpolarization-activated inward current (B, right panel) Oxytocin increased the mean peak current density at 0 mV by inducing outward potassium current in small to medium-sized DRG neurons (n=20). (C) 20 µM of atosiban (n=24) and (D) 5 µM of V1aRA blocked oxytocin-induced the voltage-activated outward potassium current and the hyperpolarization-activated inward current. TTX (0.5 µM) were treated in solution of all experiments (n=13). * p<0.01, ** p<0.001, repeated measured-ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. OT, Oxytocin; V1aRA, vasopressin-1a receptor antagonist; ns, not significant.

    Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

    Article Title: Oxytocin produces thermal analgesia via vasopressin-1a receptor by modulating TRPV1 and potassium conductance in the dorsal root ganglion neurons

    doi: 10.4196/kjpp.2018.22.2.173

    Figure Lengend Snippet: (A) Depolarizing (from –110 mV to 0 mV) ramp protocol for voltage-clamp experiments (B, left panel) Representative trace showing effects of oxytocin (5 µM) on I-V relationship response to ramp depolarization. (B, middle panel) Representative trace illustrating markedly enhanced a voltage-activated outward current and slightly enhanced a hyperpolarization-activated inward current (B, right panel) Oxytocin increased the mean peak current density at 0 mV by inducing outward potassium current in small to medium-sized DRG neurons (n=20). (C) 20 µM of atosiban (n=24) and (D) 5 µM of V1aRA blocked oxytocin-induced the voltage-activated outward potassium current and the hyperpolarization-activated inward current. TTX (0.5 µM) were treated in solution of all experiments (n=13). * p<0.01, ** p<0.001, repeated measured-ANOVA followed by posthoc Bonferroni's test. Error bars represent SEM. OT, Oxytocin; V1aRA, vasopressin-1a receptor antagonist; ns, not significant.

    Article Snippet: The primary antibodies were rabbit anti-V1a receptor polyclonal antibody (1:400; Alomone Labs, UK) and guinea pig anti-TRPV1 polyclonal antibody (1:500; Novus biologicals, USA), diluted in PBS containing 0.3% Triton X-100, 1% normal donkey serum (Jackson ImmunoResearch Co, USA), incubated overnight at 4℃ then incubated for 1 h with donkey anti-guinea pig IgG conjugated with cyanine dye 3 (Cy3, 1:200; Jackson ImmunoResearch Co, USA) and donkey anti-rabbit IgG conjugated with fluorescein-isothiocyanate (FITC, 1:200; Jackson ImmunoResearch Co, USA) diluted in the same buffer as the primary antibody.

    Techniques:

    a Serial coronal sections showing the ERα immunolabelling at the Bregma rostro-caudal coordinates (numbers in mm under the photomicrographs). Boxed area in A at higher magnification showing the exclusive nuclear labeling pattern. The bold numbered levels (A 1 and A 8 ), were chosen to show that no positive labeling was found in either rostral or caudal directions. A 9 : a horizontal view of the distribution of estrogen-receptive cells was symbolized by the red oval. A 10 : sagittal view of rat brain atlas, modified from Paxinos & Watson , at lat. 0.90 mm, where lateral habenula (LHb) is symbolized with a gray shade and A9 plane was symbolized with a horizontal line. b In situ hybridization using multiple RNAscope methods. B 1 : multiplex fluorescence method, Esr1, gene that encodes ERα (red punctuated labeling) co-expressed with Slc32a1, gene that encodes VGAT (green punctuated labeling); arrows indicate the double-labeled cells; B 2 : duplex method, Esr1 (red punctuated labeling) co-localization with Slc17a6, gene that encodes VGLUT2 (green punctuated labeling); arrows indicate the double-labeled cells; B 3 : with duplex method, Slc32a1 encoding VGAT (red punctuated labeling) shows complete co-localization with Slc17a6 encoding VGLUT2 (green punctuated labeling); arrows indicate the double-labeled cells. Inset of B 3 , Slc32a1 expression in a sexually active (SA) rat LHb, Br. −3.72 mm (brown labeling, single chromogenic-Brown method-RNAscope). *Note the similarity with ERα expression in A . c Indirect immunohistochemistry showing the GABAergic nature of the ERα+ neurons (red) in a SA rat brain. The GABA antibody (green, Sigma, A0310) produced characteristic surface labeling (C 1 , C 2 : the strip-like image was produced by Vibratome slicing leaving the brain section with an uneven surface). d ERα+ neurons co-express receptor/receptor subtypes for vasopressin, orexin, dopamine, and serotonin. D 1 : In situ hybridization using RNAscope-multiplex method targeting Esr1 (red dots), Slc32a1 (white dots) and Hcrtr2, gene encodes the orexin receptor 2 (green dots). D 2 , D 3 : Indirect immunofluorescence reactions, showing that the ERα-IR cells co-expressed vasopressin receptor V1a (inset showing the V1a antibody labeling pattern in temporal hippocampus CA2 cells body layer. See also SI Fig. for more information about this antibody), dopamine receptor D5R (also called D1Rb) and serotonin receptor 5-HTr2a, respectively. Scale bars: A 5 and b : 500 µm and rest: 10 µm

    Journal: Translational Psychiatry

    Article Title: A GABAergic cell type in the lateral habenula links hypothalamic homeostatic and midbrain motivation circuits with sex steroid signaling

    doi: 10.1038/s41398-018-0099-5

    Figure Lengend Snippet: a Serial coronal sections showing the ERα immunolabelling at the Bregma rostro-caudal coordinates (numbers in mm under the photomicrographs). Boxed area in A at higher magnification showing the exclusive nuclear labeling pattern. The bold numbered levels (A 1 and A 8 ), were chosen to show that no positive labeling was found in either rostral or caudal directions. A 9 : a horizontal view of the distribution of estrogen-receptive cells was symbolized by the red oval. A 10 : sagittal view of rat brain atlas, modified from Paxinos & Watson , at lat. 0.90 mm, where lateral habenula (LHb) is symbolized with a gray shade and A9 plane was symbolized with a horizontal line. b In situ hybridization using multiple RNAscope methods. B 1 : multiplex fluorescence method, Esr1, gene that encodes ERα (red punctuated labeling) co-expressed with Slc32a1, gene that encodes VGAT (green punctuated labeling); arrows indicate the double-labeled cells; B 2 : duplex method, Esr1 (red punctuated labeling) co-localization with Slc17a6, gene that encodes VGLUT2 (green punctuated labeling); arrows indicate the double-labeled cells; B 3 : with duplex method, Slc32a1 encoding VGAT (red punctuated labeling) shows complete co-localization with Slc17a6 encoding VGLUT2 (green punctuated labeling); arrows indicate the double-labeled cells. Inset of B 3 , Slc32a1 expression in a sexually active (SA) rat LHb, Br. −3.72 mm (brown labeling, single chromogenic-Brown method-RNAscope). *Note the similarity with ERα expression in A . c Indirect immunohistochemistry showing the GABAergic nature of the ERα+ neurons (red) in a SA rat brain. The GABA antibody (green, Sigma, A0310) produced characteristic surface labeling (C 1 , C 2 : the strip-like image was produced by Vibratome slicing leaving the brain section with an uneven surface). d ERα+ neurons co-express receptor/receptor subtypes for vasopressin, orexin, dopamine, and serotonin. D 1 : In situ hybridization using RNAscope-multiplex method targeting Esr1 (red dots), Slc32a1 (white dots) and Hcrtr2, gene encodes the orexin receptor 2 (green dots). D 2 , D 3 : Indirect immunofluorescence reactions, showing that the ERα-IR cells co-expressed vasopressin receptor V1a (inset showing the V1a antibody labeling pattern in temporal hippocampus CA2 cells body layer. See also SI Fig. for more information about this antibody), dopamine receptor D5R (also called D1Rb) and serotonin receptor 5-HTr2a, respectively. Scale bars: A 5 and b : 500 µm and rest: 10 µm

    Article Snippet: Buijs , 1:2000), tyrosine hydroxylase (sheep anti-TH, EMD Millipore Corporation, MA, AB-1542, 1:4000), serotonin transporter (goat anti-SerT, Santa Cruz Biotechnology, CA, SC-1458, 1:2000), hypocretin/orexin (rabbit anti-OR, gift from A. van del Pol ), vesicular glutamate transporter 2 (guinea pig anti-VGluT2, Frontier Institute, Co., Japan, gp-AF240-1, 1:1000), vesicular inhibitory amino acid transporter (rabbit anti-VGAT/VIAAT, provided by L. E. Eiden , 1:1000), GABA (mouse anti-GABA, Sigma-Aldrich Co. MO, A0310, 1:1000), glutamic acid decarboxylase 65 kDa isoform (mouse anti-GAD-65, EMD Millipore Co. MA, MAB351, 1:2000), glutamic acid decarboxylase 67 kDa isoform (mouse anti-GAD-67, EMD Millipore Co. MA, MAB5406, 1:2000), parvalbumin (mouse anti-PV, Swant, Switzerland, Cat. 235, 1:5000), P450 Aromatase (rabbit anti-ARO, provided by L. M. García-Segura , 1:2000), P450 Aromatase (rabbit anti-ARO, Abcam, Cambrdge, UK, AB18995, 1:2000), P450 Aromatase (mouse anti-ARO, Acris, SM2222P, 1:200), estrogen receptor-alpha (rabbit anti-ERα, Santa Cruz, CA, SC542, 1:2000), androgen receptor (rabbit anti-AR, Santa Cruz, CA, SC816, 1:2000), dopamine receptor 5 (rabbit anti-D5R, also called D1Rb, Alomone, Israel, 1:1000), serotonin receptor (mouse anti-5-HTR2a, BD pharmingen, Cat. 556326, 1:200), vasopressin receptor 1a (rabbit anti-V1a, provided by K. Mutig and T. Giesecke, see and SI Fig. for details), and green fluorescent protein (mouse anti-GFP, Abcam, Cambridge, UK, Ab291-50, 1:500).

    Techniques: Labeling, Modification, In Situ Hybridization, Multiplex Assay, Fluorescence, Expressing, Immunohistochemistry, Produced, Stripping Membranes, Immunofluorescence, Antibody Labeling