rabbit polyclonal anti gaba a α4 receptor α4  (Alomone Labs)


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    Alomone Labs rabbit polyclonal anti gaba a α4 receptor α4
    Rabbit Polyclonal Anti Gaba A α4 Receptor α4, 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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    rabbit polyclonal anti gaba a α4 receptor α4  (Alomone Labs)


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    Alomone Labs rabbit polyclonal anti gaba a α4 receptor α4
    Rabbit Polyclonal Anti Gaba A α4 Receptor α4, 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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    aga  (Alomone Labs)


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    Alomone Labs aga
    Aga, 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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    rabbit polyclonal anti gaba a α4 receptor  (Alomone Labs)


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    Alomone Labs rabbit polyclonal anti gaba a α4 receptor
    (A) Shisa7 interacted with α5 in HEK293T cells. HEK293T cells were transfected with Flag-Shisa7, α5-GFP, or both Flag-Shisa7 and α5-GFP. Cell lysates were subjected to immunoprecipitation and immunoblotting assays (n = 3 independent experiments). (B) α5, but not <t>α4</t> or δ, subunit was co-immunoprecipitated with Shisa7 in detergent-solubilized mouse hippocampal lysates. Asterisk indicates the Shisa7 band (n = 3 independent experiments). (C) Co-expression of α5β3γ2 with Shisa7-IRES-GFP (Shisa7/GFP), but not GFP, increased surface levels, total levels, and surface to total ratio of α5 expression (GFP, n = 23; Shisa7/GFP, n = 27; surface α5: t test, p < 0.0001; total α5: Mann-Whitney U test, p = 0.0298; surface α5/ total α5: t test, p = 0.0009). (D) GABA-evoked α5β3γ2-mediated whole-cell currents were significantly increased in HEK293T cells co-expressing Shisa7/GFP. (GFP, n = 17; Shisa7/GFP, n = 14, t test, p = 0.0001). (E) Immunostaining (left) and summary graphs (right) showing surface and total α5 expression were reduced in Shisa7 KO neurons (WT, n = 22; KO, n = 20; surface α5: Mann-Whitney U test, p < 0.0001; total α5: Mann-Whitney U test, p < 0.0001; surface α5/total α5: Mann-Whitney U test, p < 0.0001). (F) Representative images (left) of SEP-α5 fluorescence and the regions of the neuronal dendrites used for the fluorescence recovery after photobleaching (FRAP) and fluorescence loss in photobleaching (FLIP) experiments. Repetitive photobleaching (FLIP, white box) occurred at regions bilateral to the central FRAP region (red box). Each column (right) represents before (pre), immediately after (t = 0′), and at 2 min (t = 2′) and 5 min (t = 5′) after photobleaching in each condition. (G) Normalized fluorescence recovery curves showing significantly less newly inserted α5 on the cell surface in Shisa7 KO neurons than in WT neurons 5 min after photobleaching, indicating that α5-GABA A Rs exocytosis was impaired inShisa7 KO neurons (WT, n = 6; KO, n = 8, two-way ANOVA with Tukey’s multiple comparison test. t = 5′: p < 0.0001; t = 5.5′: p = 0.0002; t = 6′: p < 0.0001; t = 6.5′: p = 0.0002). *p < 0.05, ***p < 0.001, and ****p < 0.0001. All data are presented as mean ± SEM. See also .
    Rabbit Polyclonal Anti Gaba A α4 Receptor, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti gaba a α4 receptor/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
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    rabbit polyclonal anti gaba a α4 receptor - by Bioz Stars, 2023-02
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    1) Product Images from "Activity- and sleep-dependent regulation of tonic inhibition by Shisa7"

    Article Title: Activity- and sleep-dependent regulation of tonic inhibition by Shisa7

    Journal: Cell reports

    doi: 10.1016/j.celrep.2021.108899

    (A) Shisa7 interacted with α5 in HEK293T cells. HEK293T cells were transfected with Flag-Shisa7, α5-GFP, or both Flag-Shisa7 and α5-GFP. Cell lysates were subjected to immunoprecipitation and immunoblotting assays (n = 3 independent experiments). (B) α5, but not α4 or δ, subunit was co-immunoprecipitated with Shisa7 in detergent-solubilized mouse hippocampal lysates. Asterisk indicates the Shisa7 band (n = 3 independent experiments). (C) Co-expression of α5β3γ2 with Shisa7-IRES-GFP (Shisa7/GFP), but not GFP, increased surface levels, total levels, and surface to total ratio of α5 expression (GFP, n = 23; Shisa7/GFP, n = 27; surface α5: t test, p < 0.0001; total α5: Mann-Whitney U test, p = 0.0298; surface α5/ total α5: t test, p = 0.0009). (D) GABA-evoked α5β3γ2-mediated whole-cell currents were significantly increased in HEK293T cells co-expressing Shisa7/GFP. (GFP, n = 17; Shisa7/GFP, n = 14, t test, p = 0.0001). (E) Immunostaining (left) and summary graphs (right) showing surface and total α5 expression were reduced in Shisa7 KO neurons (WT, n = 22; KO, n = 20; surface α5: Mann-Whitney U test, p < 0.0001; total α5: Mann-Whitney U test, p < 0.0001; surface α5/total α5: Mann-Whitney U test, p < 0.0001). (F) Representative images (left) of SEP-α5 fluorescence and the regions of the neuronal dendrites used for the fluorescence recovery after photobleaching (FRAP) and fluorescence loss in photobleaching (FLIP) experiments. Repetitive photobleaching (FLIP, white box) occurred at regions bilateral to the central FRAP region (red box). Each column (right) represents before (pre), immediately after (t = 0′), and at 2 min (t = 2′) and 5 min (t = 5′) after photobleaching in each condition. (G) Normalized fluorescence recovery curves showing significantly less newly inserted α5 on the cell surface in Shisa7 KO neurons than in WT neurons 5 min after photobleaching, indicating that α5-GABA A Rs exocytosis was impaired inShisa7 KO neurons (WT, n = 6; KO, n = 8, two-way ANOVA with Tukey’s multiple comparison test. t = 5′: p < 0.0001; t = 5.5′: p = 0.0002; t = 6′: p < 0.0001; t = 6.5′: p = 0.0002). *p < 0.05, ***p < 0.001, and ****p < 0.0001. All data are presented as mean ± SEM. See also .
    Figure Legend Snippet: (A) Shisa7 interacted with α5 in HEK293T cells. HEK293T cells were transfected with Flag-Shisa7, α5-GFP, or both Flag-Shisa7 and α5-GFP. Cell lysates were subjected to immunoprecipitation and immunoblotting assays (n = 3 independent experiments). (B) α5, but not α4 or δ, subunit was co-immunoprecipitated with Shisa7 in detergent-solubilized mouse hippocampal lysates. Asterisk indicates the Shisa7 band (n = 3 independent experiments). (C) Co-expression of α5β3γ2 with Shisa7-IRES-GFP (Shisa7/GFP), but not GFP, increased surface levels, total levels, and surface to total ratio of α5 expression (GFP, n = 23; Shisa7/GFP, n = 27; surface α5: t test, p < 0.0001; total α5: Mann-Whitney U test, p = 0.0298; surface α5/ total α5: t test, p = 0.0009). (D) GABA-evoked α5β3γ2-mediated whole-cell currents were significantly increased in HEK293T cells co-expressing Shisa7/GFP. (GFP, n = 17; Shisa7/GFP, n = 14, t test, p = 0.0001). (E) Immunostaining (left) and summary graphs (right) showing surface and total α5 expression were reduced in Shisa7 KO neurons (WT, n = 22; KO, n = 20; surface α5: Mann-Whitney U test, p < 0.0001; total α5: Mann-Whitney U test, p < 0.0001; surface α5/total α5: Mann-Whitney U test, p < 0.0001). (F) Representative images (left) of SEP-α5 fluorescence and the regions of the neuronal dendrites used for the fluorescence recovery after photobleaching (FRAP) and fluorescence loss in photobleaching (FLIP) experiments. Repetitive photobleaching (FLIP, white box) occurred at regions bilateral to the central FRAP region (red box). Each column (right) represents before (pre), immediately after (t = 0′), and at 2 min (t = 2′) and 5 min (t = 5′) after photobleaching in each condition. (G) Normalized fluorescence recovery curves showing significantly less newly inserted α5 on the cell surface in Shisa7 KO neurons than in WT neurons 5 min after photobleaching, indicating that α5-GABA A Rs exocytosis was impaired inShisa7 KO neurons (WT, n = 6; KO, n = 8, two-way ANOVA with Tukey’s multiple comparison test. t = 5′: p < 0.0001; t = 5.5′: p = 0.0002; t = 6′: p < 0.0001; t = 6.5′: p = 0.0002). *p < 0.05, ***p < 0.001, and ****p < 0.0001. All data are presented as mean ± SEM. See also .

    Techniques Used: Transfection, Immunoprecipitation, Western Blot, Expressing, MANN-WHITNEY, Immunostaining, Fluorescence

    KEY RESOURCES TABLE
    Figure Legend Snippet: KEY RESOURCES TABLE

    Techniques Used: Recombinant, Transfection, Protease Inhibitor, Knock-Out, Software

    aga 008  (Alomone Labs)


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    Alomone Labs aga 008
    Aga 008, 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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    gaba a α4  (Alomone Labs)


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    Alomone Labs gaba a α4
    Primary antibodies
    Gaba A α4, 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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    1) Product Images from "Tonic GABAergic inhibition, via GABA A receptors containing αβƐ subunits, regulates excitability of ventral tegmental area dopamine neurons"

    Article Title: Tonic GABAergic inhibition, via GABA A receptors containing αβƐ subunits, regulates excitability of ventral tegmental area dopamine neurons

    Journal: The European Journal of Neuroscience

    doi: 10.1111/ejn.15133

    Primary antibodies
    Figure Legend Snippet: Primary antibodies

    Techniques Used:

    rabbit anti α4 gaba a r  (Alomone Labs)


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    Alomone Labs rabbit anti α4 gaba a r
    (a) Southern blot screen of ES cell lines: the Q241M mutation silently introduces a novel restriction site – allowing a screen for restriction fragment length polymorphism. Lanes 1 - 5, heterozygous recombinants; lanes 6 - 7, wild-type littermate controls. (b) Verifying the point mutation by DNA sequencing. Exon 8 was PCR-amplified from mouse genomic DNA of wild-type (α2 Q/Q : left), α2 Q/M (middle) and α2 M/M (right) animals, then sequenced. Sequence data around the point mutation are shown. (c,d) <t>GABA</t> <t>A</t> <t>R</t> <t>α1-α4</t> subunit expression levels were determined by Western blotting total protein isolated from the cortex ( c ) and hippocampus ( d ). Left panels, quantitation of expression in α2 Q/M (grey) and α2 M/M (white) relative to wild-type (α2 Q/Q , black) brain samples. Right panels, representative Western blots from cortex and hippocampus for wild-type (WT), α2 Q/M and α2 M/M mice. Images show blots between 45 and 66 kDa markers. Loading controls with tubulin are omitted for clarity. Data are mean ± sem, one-way ANOVA, n = 6-9. There are no statistically significant differences between groups.
    Rabbit Anti α4 Gaba A R, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti α4 gaba a r/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
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    rabbit anti α4 gaba a r - by Bioz Stars, 2023-02
    94/100 stars

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    1) Product Images from "Brain neurosteroids are natural anxiolytics targeting α2 subunit γ-aminobutyric acid type-A receptors"

    Article Title: Brain neurosteroids are natural anxiolytics targeting α2 subunit γ-aminobutyric acid type-A receptors

    Journal: bioRxiv

    doi: 10.1101/462457

    (a) Southern blot screen of ES cell lines: the Q241M mutation silently introduces a novel restriction site – allowing a screen for restriction fragment length polymorphism. Lanes 1 - 5, heterozygous recombinants; lanes 6 - 7, wild-type littermate controls. (b) Verifying the point mutation by DNA sequencing. Exon 8 was PCR-amplified from mouse genomic DNA of wild-type (α2 Q/Q : left), α2 Q/M (middle) and α2 M/M (right) animals, then sequenced. Sequence data around the point mutation are shown. (c,d) GABA A R α1-α4 subunit expression levels were determined by Western blotting total protein isolated from the cortex ( c ) and hippocampus ( d ). Left panels, quantitation of expression in α2 Q/M (grey) and α2 M/M (white) relative to wild-type (α2 Q/Q , black) brain samples. Right panels, representative Western blots from cortex and hippocampus for wild-type (WT), α2 Q/M and α2 M/M mice. Images show blots between 45 and 66 kDa markers. Loading controls with tubulin are omitted for clarity. Data are mean ± sem, one-way ANOVA, n = 6-9. There are no statistically significant differences between groups.
    Figure Legend Snippet: (a) Southern blot screen of ES cell lines: the Q241M mutation silently introduces a novel restriction site – allowing a screen for restriction fragment length polymorphism. Lanes 1 - 5, heterozygous recombinants; lanes 6 - 7, wild-type littermate controls. (b) Verifying the point mutation by DNA sequencing. Exon 8 was PCR-amplified from mouse genomic DNA of wild-type (α2 Q/Q : left), α2 Q/M (middle) and α2 M/M (right) animals, then sequenced. Sequence data around the point mutation are shown. (c,d) GABA A R α1-α4 subunit expression levels were determined by Western blotting total protein isolated from the cortex ( c ) and hippocampus ( d ). Left panels, quantitation of expression in α2 Q/M (grey) and α2 M/M (white) relative to wild-type (α2 Q/Q , black) brain samples. Right panels, representative Western blots from cortex and hippocampus for wild-type (WT), α2 Q/M and α2 M/M mice. Images show blots between 45 and 66 kDa markers. Loading controls with tubulin are omitted for clarity. Data are mean ± sem, one-way ANOVA, n = 6-9. There are no statistically significant differences between groups.

    Techniques Used: Southern Blot, Mutagenesis, DNA Sequencing, Amplification, Sequencing, Expressing, Western Blot, Isolation, Quantitation Assay

    rabbit anti α4 gaba a r  (Alomone Labs)


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    Alomone Labs rabbit anti α4 gaba a r
    (a) Southern blot screen of ES cell lines: the Q241M mutation silently introduces a novel restriction site – allowing a screen for restriction fragment length polymorphism. Lanes 1 - 5, heterozygous recombinants; lanes 6 - 7, wild-type littermate controls. (b) Verifying the point mutation by DNA sequencing. Exon 8 was PCR-amplified from mouse genomic DNA of wild-type (α2 Q/Q : left), α2 Q/M (middle) and α2 M/M (right) animals, then sequenced. Sequence data around the point mutation are shown. (c,d) <t>GABA</t> <t>A</t> <t>R</t> <t>α1-α4</t> subunit expression levels were determined by Western blotting total protein isolated from the cortex ( c ) and hippocampus ( d ). Left panels, quantitation of expression in α2 Q/M (grey) and α2 M/M (white) relative to wild-type (α2 Q/Q , black) brain samples. Right panels, representative Western blots from cortex and hippocampus for wild-type (WT), α2 Q/M and α2 M/M mice. Images show blots between 45 and 66 kDa markers. Loading controls with tubulin are omitted for clarity. Data are mean ± sem, one-way ANOVA, n = 6-9. There are no statistically significant differences between groups.
    Rabbit Anti α4 Gaba A R, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti α4 gaba a r/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
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    rabbit anti α4 gaba a r - by Bioz Stars, 2023-02
    94/100 stars

    Images

    1) Product Images from "Brain neurosteroids are natural anxiolytics targeting α2 subunit γ-aminobutyric acid type-A receptors"

    Article Title: Brain neurosteroids are natural anxiolytics targeting α2 subunit γ-aminobutyric acid type-A receptors

    Journal: bioRxiv

    doi: 10.1101/462457

    (a) Southern blot screen of ES cell lines: the Q241M mutation silently introduces a novel restriction site – allowing a screen for restriction fragment length polymorphism. Lanes 1 - 5, heterozygous recombinants; lanes 6 - 7, wild-type littermate controls. (b) Verifying the point mutation by DNA sequencing. Exon 8 was PCR-amplified from mouse genomic DNA of wild-type (α2 Q/Q : left), α2 Q/M (middle) and α2 M/M (right) animals, then sequenced. Sequence data around the point mutation are shown. (c,d) GABA A R α1-α4 subunit expression levels were determined by Western blotting total protein isolated from the cortex ( c ) and hippocampus ( d ). Left panels, quantitation of expression in α2 Q/M (grey) and α2 M/M (white) relative to wild-type (α2 Q/Q , black) brain samples. Right panels, representative Western blots from cortex and hippocampus for wild-type (WT), α2 Q/M and α2 M/M mice. Images show blots between 45 and 66 kDa markers. Loading controls with tubulin are omitted for clarity. Data are mean ± sem, one-way ANOVA, n = 6-9. There are no statistically significant differences between groups.
    Figure Legend Snippet: (a) Southern blot screen of ES cell lines: the Q241M mutation silently introduces a novel restriction site – allowing a screen for restriction fragment length polymorphism. Lanes 1 - 5, heterozygous recombinants; lanes 6 - 7, wild-type littermate controls. (b) Verifying the point mutation by DNA sequencing. Exon 8 was PCR-amplified from mouse genomic DNA of wild-type (α2 Q/Q : left), α2 Q/M (middle) and α2 M/M (right) animals, then sequenced. Sequence data around the point mutation are shown. (c,d) GABA A R α1-α4 subunit expression levels were determined by Western blotting total protein isolated from the cortex ( c ) and hippocampus ( d ). Left panels, quantitation of expression in α2 Q/M (grey) and α2 M/M (white) relative to wild-type (α2 Q/Q , black) brain samples. Right panels, representative Western blots from cortex and hippocampus for wild-type (WT), α2 Q/M and α2 M/M mice. Images show blots between 45 and 66 kDa markers. Loading controls with tubulin are omitted for clarity. Data are mean ± sem, one-way ANOVA, n = 6-9. There are no statistically significant differences between groups.

    Techniques Used: Southern Blot, Mutagenesis, DNA Sequencing, Amplification, Sequencing, Expressing, Western Blot, Isolation, Quantitation Assay

    rabbit polyclonal gabaar a4 subunit antibody  (Alomone Labs)


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    Alomone Labs rabbit polyclonal gabaar a4 subunit antibody
    Rabbit Polyclonal Gabaar A4 Subunit Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal gabaar a4 subunit antibody/product/Alomone Labs
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    Alomone Labs rabbit polyclonal anti gaba a α4 receptor α4
    Rabbit Polyclonal Anti Gaba A α4 Receptor α4, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti gaba a α4 receptor α4/product/Alomone Labs
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    Alomone Labs aga
    Aga, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/aga/product/Alomone Labs
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    Alomone Labs rabbit polyclonal anti gaba a α4 receptor
    (A) Shisa7 interacted with α5 in HEK293T cells. HEK293T cells were transfected with Flag-Shisa7, α5-GFP, or both Flag-Shisa7 and α5-GFP. Cell lysates were subjected to immunoprecipitation and immunoblotting assays (n = 3 independent experiments). (B) α5, but not <t>α4</t> or δ, subunit was co-immunoprecipitated with Shisa7 in detergent-solubilized mouse hippocampal lysates. Asterisk indicates the Shisa7 band (n = 3 independent experiments). (C) Co-expression of α5β3γ2 with Shisa7-IRES-GFP (Shisa7/GFP), but not GFP, increased surface levels, total levels, and surface to total ratio of α5 expression (GFP, n = 23; Shisa7/GFP, n = 27; surface α5: t test, p < 0.0001; total α5: Mann-Whitney U test, p = 0.0298; surface α5/ total α5: t test, p = 0.0009). (D) GABA-evoked α5β3γ2-mediated whole-cell currents were significantly increased in HEK293T cells co-expressing Shisa7/GFP. (GFP, n = 17; Shisa7/GFP, n = 14, t test, p = 0.0001). (E) Immunostaining (left) and summary graphs (right) showing surface and total α5 expression were reduced in Shisa7 KO neurons (WT, n = 22; KO, n = 20; surface α5: Mann-Whitney U test, p < 0.0001; total α5: Mann-Whitney U test, p < 0.0001; surface α5/total α5: Mann-Whitney U test, p < 0.0001). (F) Representative images (left) of SEP-α5 fluorescence and the regions of the neuronal dendrites used for the fluorescence recovery after photobleaching (FRAP) and fluorescence loss in photobleaching (FLIP) experiments. Repetitive photobleaching (FLIP, white box) occurred at regions bilateral to the central FRAP region (red box). Each column (right) represents before (pre), immediately after (t = 0′), and at 2 min (t = 2′) and 5 min (t = 5′) after photobleaching in each condition. (G) Normalized fluorescence recovery curves showing significantly less newly inserted α5 on the cell surface in Shisa7 KO neurons than in WT neurons 5 min after photobleaching, indicating that α5-GABA A Rs exocytosis was impaired inShisa7 KO neurons (WT, n = 6; KO, n = 8, two-way ANOVA with Tukey’s multiple comparison test. t = 5′: p < 0.0001; t = 5.5′: p = 0.0002; t = 6′: p < 0.0001; t = 6.5′: p = 0.0002). *p < 0.05, ***p < 0.001, and ****p < 0.0001. All data are presented as mean ± SEM. See also .
    Rabbit Polyclonal Anti Gaba A α4 Receptor, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti gaba a α4 receptor/product/Alomone Labs
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    Alomone Labs aga 008
    (A) Shisa7 interacted with α5 in HEK293T cells. HEK293T cells were transfected with Flag-Shisa7, α5-GFP, or both Flag-Shisa7 and α5-GFP. Cell lysates were subjected to immunoprecipitation and immunoblotting assays (n = 3 independent experiments). (B) α5, but not <t>α4</t> or δ, subunit was co-immunoprecipitated with Shisa7 in detergent-solubilized mouse hippocampal lysates. Asterisk indicates the Shisa7 band (n = 3 independent experiments). (C) Co-expression of α5β3γ2 with Shisa7-IRES-GFP (Shisa7/GFP), but not GFP, increased surface levels, total levels, and surface to total ratio of α5 expression (GFP, n = 23; Shisa7/GFP, n = 27; surface α5: t test, p < 0.0001; total α5: Mann-Whitney U test, p = 0.0298; surface α5/ total α5: t test, p = 0.0009). (D) GABA-evoked α5β3γ2-mediated whole-cell currents were significantly increased in HEK293T cells co-expressing Shisa7/GFP. (GFP, n = 17; Shisa7/GFP, n = 14, t test, p = 0.0001). (E) Immunostaining (left) and summary graphs (right) showing surface and total α5 expression were reduced in Shisa7 KO neurons (WT, n = 22; KO, n = 20; surface α5: Mann-Whitney U test, p < 0.0001; total α5: Mann-Whitney U test, p < 0.0001; surface α5/total α5: Mann-Whitney U test, p < 0.0001). (F) Representative images (left) of SEP-α5 fluorescence and the regions of the neuronal dendrites used for the fluorescence recovery after photobleaching (FRAP) and fluorescence loss in photobleaching (FLIP) experiments. Repetitive photobleaching (FLIP, white box) occurred at regions bilateral to the central FRAP region (red box). Each column (right) represents before (pre), immediately after (t = 0′), and at 2 min (t = 2′) and 5 min (t = 5′) after photobleaching in each condition. (G) Normalized fluorescence recovery curves showing significantly less newly inserted α5 on the cell surface in Shisa7 KO neurons than in WT neurons 5 min after photobleaching, indicating that α5-GABA A Rs exocytosis was impaired inShisa7 KO neurons (WT, n = 6; KO, n = 8, two-way ANOVA with Tukey’s multiple comparison test. t = 5′: p < 0.0001; t = 5.5′: p = 0.0002; t = 6′: p < 0.0001; t = 6.5′: p = 0.0002). *p < 0.05, ***p < 0.001, and ****p < 0.0001. All data are presented as mean ± SEM. See also .
    Aga 008, 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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    Alomone Labs gaba a α4
    Primary antibodies
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    Alomone Labs rabbit anti α4 gaba a r
    (a) Southern blot screen of ES cell lines: the Q241M mutation silently introduces a novel restriction site – allowing a screen for restriction fragment length polymorphism. Lanes 1 - 5, heterozygous recombinants; lanes 6 - 7, wild-type littermate controls. (b) Verifying the point mutation by DNA sequencing. Exon 8 was PCR-amplified from mouse genomic DNA of wild-type (α2 Q/Q : left), α2 Q/M (middle) and α2 M/M (right) animals, then sequenced. Sequence data around the point mutation are shown. (c,d) <t>GABA</t> <t>A</t> <t>R</t> <t>α1-α4</t> subunit expression levels were determined by Western blotting total protein isolated from the cortex ( c ) and hippocampus ( d ). Left panels, quantitation of expression in α2 Q/M (grey) and α2 M/M (white) relative to wild-type (α2 Q/Q , black) brain samples. Right panels, representative Western blots from cortex and hippocampus for wild-type (WT), α2 Q/M and α2 M/M mice. Images show blots between 45 and 66 kDa markers. Loading controls with tubulin are omitted for clarity. Data are mean ± sem, one-way ANOVA, n = 6-9. There are no statistically significant differences between groups.
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    Alomone Labs rabbit polyclonal gabaar a4 subunit antibody
    (a) Southern blot screen of ES cell lines: the Q241M mutation silently introduces a novel restriction site – allowing a screen for restriction fragment length polymorphism. Lanes 1 - 5, heterozygous recombinants; lanes 6 - 7, wild-type littermate controls. (b) Verifying the point mutation by DNA sequencing. Exon 8 was PCR-amplified from mouse genomic DNA of wild-type (α2 Q/Q : left), α2 Q/M (middle) and α2 M/M (right) animals, then sequenced. Sequence data around the point mutation are shown. (c,d) <t>GABA</t> <t>A</t> <t>R</t> <t>α1-α4</t> subunit expression levels were determined by Western blotting total protein isolated from the cortex ( c ) and hippocampus ( d ). Left panels, quantitation of expression in α2 Q/M (grey) and α2 M/M (white) relative to wild-type (α2 Q/Q , black) brain samples. Right panels, representative Western blots from cortex and hippocampus for wild-type (WT), α2 Q/M and α2 M/M mice. Images show blots between 45 and 66 kDa markers. Loading controls with tubulin are omitted for clarity. Data are mean ± sem, one-way ANOVA, n = 6-9. There are no statistically significant differences between groups.
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    Image Search Results


    (A) Shisa7 interacted with α5 in HEK293T cells. HEK293T cells were transfected with Flag-Shisa7, α5-GFP, or both Flag-Shisa7 and α5-GFP. Cell lysates were subjected to immunoprecipitation and immunoblotting assays (n = 3 independent experiments). (B) α5, but not α4 or δ, subunit was co-immunoprecipitated with Shisa7 in detergent-solubilized mouse hippocampal lysates. Asterisk indicates the Shisa7 band (n = 3 independent experiments). (C) Co-expression of α5β3γ2 with Shisa7-IRES-GFP (Shisa7/GFP), but not GFP, increased surface levels, total levels, and surface to total ratio of α5 expression (GFP, n = 23; Shisa7/GFP, n = 27; surface α5: t test, p < 0.0001; total α5: Mann-Whitney U test, p = 0.0298; surface α5/ total α5: t test, p = 0.0009). (D) GABA-evoked α5β3γ2-mediated whole-cell currents were significantly increased in HEK293T cells co-expressing Shisa7/GFP. (GFP, n = 17; Shisa7/GFP, n = 14, t test, p = 0.0001). (E) Immunostaining (left) and summary graphs (right) showing surface and total α5 expression were reduced in Shisa7 KO neurons (WT, n = 22; KO, n = 20; surface α5: Mann-Whitney U test, p < 0.0001; total α5: Mann-Whitney U test, p < 0.0001; surface α5/total α5: Mann-Whitney U test, p < 0.0001). (F) Representative images (left) of SEP-α5 fluorescence and the regions of the neuronal dendrites used for the fluorescence recovery after photobleaching (FRAP) and fluorescence loss in photobleaching (FLIP) experiments. Repetitive photobleaching (FLIP, white box) occurred at regions bilateral to the central FRAP region (red box). Each column (right) represents before (pre), immediately after (t = 0′), and at 2 min (t = 2′) and 5 min (t = 5′) after photobleaching in each condition. (G) Normalized fluorescence recovery curves showing significantly less newly inserted α5 on the cell surface in Shisa7 KO neurons than in WT neurons 5 min after photobleaching, indicating that α5-GABA A Rs exocytosis was impaired inShisa7 KO neurons (WT, n = 6; KO, n = 8, two-way ANOVA with Tukey’s multiple comparison test. t = 5′: p < 0.0001; t = 5.5′: p = 0.0002; t = 6′: p < 0.0001; t = 6.5′: p = 0.0002). *p < 0.05, ***p < 0.001, and ****p < 0.0001. All data are presented as mean ± SEM. See also .

    Journal: Cell reports

    Article Title: Activity- and sleep-dependent regulation of tonic inhibition by Shisa7

    doi: 10.1016/j.celrep.2021.108899

    Figure Lengend Snippet: (A) Shisa7 interacted with α5 in HEK293T cells. HEK293T cells were transfected with Flag-Shisa7, α5-GFP, or both Flag-Shisa7 and α5-GFP. Cell lysates were subjected to immunoprecipitation and immunoblotting assays (n = 3 independent experiments). (B) α5, but not α4 or δ, subunit was co-immunoprecipitated with Shisa7 in detergent-solubilized mouse hippocampal lysates. Asterisk indicates the Shisa7 band (n = 3 independent experiments). (C) Co-expression of α5β3γ2 with Shisa7-IRES-GFP (Shisa7/GFP), but not GFP, increased surface levels, total levels, and surface to total ratio of α5 expression (GFP, n = 23; Shisa7/GFP, n = 27; surface α5: t test, p < 0.0001; total α5: Mann-Whitney U test, p = 0.0298; surface α5/ total α5: t test, p = 0.0009). (D) GABA-evoked α5β3γ2-mediated whole-cell currents were significantly increased in HEK293T cells co-expressing Shisa7/GFP. (GFP, n = 17; Shisa7/GFP, n = 14, t test, p = 0.0001). (E) Immunostaining (left) and summary graphs (right) showing surface and total α5 expression were reduced in Shisa7 KO neurons (WT, n = 22; KO, n = 20; surface α5: Mann-Whitney U test, p < 0.0001; total α5: Mann-Whitney U test, p < 0.0001; surface α5/total α5: Mann-Whitney U test, p < 0.0001). (F) Representative images (left) of SEP-α5 fluorescence and the regions of the neuronal dendrites used for the fluorescence recovery after photobleaching (FRAP) and fluorescence loss in photobleaching (FLIP) experiments. Repetitive photobleaching (FLIP, white box) occurred at regions bilateral to the central FRAP region (red box). Each column (right) represents before (pre), immediately after (t = 0′), and at 2 min (t = 2′) and 5 min (t = 5′) after photobleaching in each condition. (G) Normalized fluorescence recovery curves showing significantly less newly inserted α5 on the cell surface in Shisa7 KO neurons than in WT neurons 5 min after photobleaching, indicating that α5-GABA A Rs exocytosis was impaired inShisa7 KO neurons (WT, n = 6; KO, n = 8, two-way ANOVA with Tukey’s multiple comparison test. t = 5′: p < 0.0001; t = 5.5′: p = 0.0002; t = 6′: p < 0.0001; t = 6.5′: p = 0.0002). *p < 0.05, ***p < 0.001, and ****p < 0.0001. All data are presented as mean ± SEM. See also .

    Article Snippet: Rabbit Polyclonal Anti-GABA(A) α4 Receptor , Alomone Labs , Cat # AGA-008; RRID: AB_10917596.

    Techniques: Transfection, Immunoprecipitation, Western Blot, Expressing, MANN-WHITNEY, Immunostaining, Fluorescence

    KEY RESOURCES TABLE

    Journal: Cell reports

    Article Title: Activity- and sleep-dependent regulation of tonic inhibition by Shisa7

    doi: 10.1016/j.celrep.2021.108899

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: Rabbit Polyclonal Anti-GABA(A) α4 Receptor , Alomone Labs , Cat # AGA-008; RRID: AB_10917596.

    Techniques: Recombinant, Transfection, Protease Inhibitor, Knock-Out, Software

    Primary antibodies

    Journal: The European Journal of Neuroscience

    Article Title: Tonic GABAergic inhibition, via GABA A receptors containing αβƐ subunits, regulates excitability of ventral tegmental area dopamine neurons

    doi: 10.1111/ejn.15133

    Figure Lengend Snippet: Primary antibodies

    Article Snippet: GABA A α4 , RbP , 1:500 , Alomone Labs , AGA‐008 , AB_10917596.

    Techniques:

    (a) Southern blot screen of ES cell lines: the Q241M mutation silently introduces a novel restriction site – allowing a screen for restriction fragment length polymorphism. Lanes 1 - 5, heterozygous recombinants; lanes 6 - 7, wild-type littermate controls. (b) Verifying the point mutation by DNA sequencing. Exon 8 was PCR-amplified from mouse genomic DNA of wild-type (α2 Q/Q : left), α2 Q/M (middle) and α2 M/M (right) animals, then sequenced. Sequence data around the point mutation are shown. (c,d) GABA A R α1-α4 subunit expression levels were determined by Western blotting total protein isolated from the cortex ( c ) and hippocampus ( d ). Left panels, quantitation of expression in α2 Q/M (grey) and α2 M/M (white) relative to wild-type (α2 Q/Q , black) brain samples. Right panels, representative Western blots from cortex and hippocampus for wild-type (WT), α2 Q/M and α2 M/M mice. Images show blots between 45 and 66 kDa markers. Loading controls with tubulin are omitted for clarity. Data are mean ± sem, one-way ANOVA, n = 6-9. There are no statistically significant differences between groups.

    Journal: bioRxiv

    Article Title: Brain neurosteroids are natural anxiolytics targeting α2 subunit γ-aminobutyric acid type-A receptors

    doi: 10.1101/462457

    Figure Lengend Snippet: (a) Southern blot screen of ES cell lines: the Q241M mutation silently introduces a novel restriction site – allowing a screen for restriction fragment length polymorphism. Lanes 1 - 5, heterozygous recombinants; lanes 6 - 7, wild-type littermate controls. (b) Verifying the point mutation by DNA sequencing. Exon 8 was PCR-amplified from mouse genomic DNA of wild-type (α2 Q/Q : left), α2 Q/M (middle) and α2 M/M (right) animals, then sequenced. Sequence data around the point mutation are shown. (c,d) GABA A R α1-α4 subunit expression levels were determined by Western blotting total protein isolated from the cortex ( c ) and hippocampus ( d ). Left panels, quantitation of expression in α2 Q/M (grey) and α2 M/M (white) relative to wild-type (α2 Q/Q , black) brain samples. Right panels, representative Western blots from cortex and hippocampus for wild-type (WT), α2 Q/M and α2 M/M mice. Images show blots between 45 and 66 kDa markers. Loading controls with tubulin are omitted for clarity. Data are mean ± sem, one-way ANOVA, n = 6-9. There are no statistically significant differences between groups.

    Article Snippet: For immunohistochemistry, the following primary antibodies were used: rabbit anti-α1-GABA A R (1:20000, from Jean-Marc Fritschy, University of Zurich), guinea-pig anti-α2-GABA A R (1:1000, from Jean-Marc Fritschy), rabbit anti-α3-GABA A R (1:1000, Alomone Labs), rabbit anti-α4-GABA A R (1:500, Werner Sieghart), rabbit anti-α5-GABA A R (1:500, Werner Sieghart).

    Techniques: Southern Blot, Mutagenesis, DNA Sequencing, Amplification, Sequencing, Expressing, Western Blot, Isolation, Quantitation Assay