rabbit anti gephyrin polyclonal antibody  (Alomone Labs)

Journal: Journal of Biomedical Science

Article Title: Generational synaptic functions of GABAA receptor β3 subunit deteriorations in an animal model of social deficit

doi: 10.1186/s12929-022-00835-w

Figure Lengend Snippet: NMDA-induced gephyrin and GABA A R enhancements in two generations of the VPA-induced offspring. a–d Representative western blot profile, the protein levels ( a , c ), and the main effect plots of two-way ANOVA ( b , d ) of gephyrin and GABA A R \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\upbeta }$$\end{document} β 3 subunit in synaptoneurosomes from the amygdala. Gephyrin, n = 7 rats from three or four litters for each condition; GABA A R \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\upbeta }$$\end{document} β 3 subunit: n = 9 rats from three or four litters for each condition. e Representative mIPSCs traces from each group. Scale bars, 100 pA, 50 ms. f , i Bar graphs show average mIPSC amplitudes ( f ) and frequency ( i ) measured from the BLA pyramidal neurons. g , j Cumulative probability of mIPSC amplitudes ( g ) and frequency ( j ) measured from the BLA pyramidal neurons. h , k The main effect plots of two-way ANOVA of mIPSC amplitudes ( h ) and frequency ( k ) measured from the BLA pyramidal neurons. n = 8–10 cells from 4–5 rats from three or four litters for each condition. Data are presented as mean ± SEM; *p

Article Snippet: For GABAA R insertion, N -methyl-d -aspartate (NMDA) was purchased from Tocris (Minneapolis, MN), which was dissolved in artificial cerebrospinal fluid (aCSF) to obtain a concentration of 20 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\upmu }$$\end{document} μ M. For western blotting, the following antibodies of GluN2A (1:5000; Genetex; GTX63442), GluN2B (1:5000; Abcam; ab65783), GluA1 (1:5000; Abcam; ab109450), GluA2 (1:5000; Millipore; MAB397), gephyrin (1:5000; Alomone; AIP-005), GABAA R β3 subunit (1:5000; Abcam; ab98968), and anti- \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\upbeta }$$\end{document} β-actin antibody (1:10,000; Abcam; ab6276) were used.

Techniques: Western Blot

Journal: Journal of Biomedical Science

Article Title: Generational synaptic functions of GABAA receptor β3 subunit deteriorations in an animal model of social deficit

doi: 10.1186/s12929-022-00835-w

Figure Lengend Snippet: Synaptic gephyrin levels and gephyrin-GABA A R associations in two generations of the VPA-induced offspring. a , b Representative western blot profile and the protein levels of gephyrin ( a ) and GABA A R \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\upbeta }$$\end{document} β 3 subunit ( b ) in synaptoneurosomes from the amygdala. Gephyrin, n = 8 rats from three or four litters for each condition; GABA A R \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\upbeta }$$\end{document} β 3 subunit, n = 7 rats from three or four litters for each condition. c , d Representative western blot profile and the protein levels of gephyrin ( c ) and GABA A R \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\upbeta }$$\end{document} β 3 subunit ( d ) in whole-cell lysate from the amygdala. Gephyrin, n = 6 rats from three or four litters for each condition; GABA A R \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\upbeta }$$\end{document} β 3 subunit, n = 6 rats from three or four litters for each condition. ( e ) Representative western blot profile and the protein levels of co-immunoprecipitation of gephyrin and GABA A R \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\upbeta }$$\end{document} β 3 subunit in synaptoneurosomes from the amygdala. n = 6 rats from three or four litters for each condition. Data are presented as mean ± SEM; *p

Article Snippet: For GABAA R insertion, N -methyl-d -aspartate (NMDA) was purchased from Tocris (Minneapolis, MN), which was dissolved in artificial cerebrospinal fluid (aCSF) to obtain a concentration of 20 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\upmu }$$\end{document} μ M. For western blotting, the following antibodies of GluN2A (1:5000; Genetex; GTX63442), GluN2B (1:5000; Abcam; ab65783), GluA1 (1:5000; Abcam; ab109450), GluA2 (1:5000; Millipore; MAB397), gephyrin (1:5000; Alomone; AIP-005), GABAA R β3 subunit (1:5000; Abcam; ab98968), and anti- \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\upbeta }$$\end{document} β-actin antibody (1:10,000; Abcam; ab6276) were used.

Techniques: Western Blot, Immunoprecipitation

Journal: BMC Neuroscience

Article Title: Glycine receptors expression in rat spinal cord and dorsal root ganglion in prostaglandin E2 intrathecal injection models

doi: 10.1186/s12868-018-0470-8

Figure Lengend Snippet: a Western blot and quantitative analysis of b gephyrin, c GlyRα3 expressions in L5 spinal cord dorsal horn at 1 h and 5 h after PGE2 intrathecal injection. The GlyRα3 protein expressions were significantly decreased in the PGE2 group at 1 h and lasted up to 5 h. The gephyrin expression decreased significantly at 5 h after PGE2 injection. d Confocal microscopic imaging showed expression of gephyrin throughout the gray matter of spinal cord. e Expression of GlyRα3 was mainly in the superficial layer of dorsal horn and in the ventral horn. Each group had n = 3–6 rats. Scale Bars: 200 μm; ** p

Article Snippet: The membranes were firstly blocked with 5% milk in phosphate-buffered saline (PBS) with 0.1% Tween-20 for 1 h at room temperature, and then probed overnight at 4 °C with Rabbit anti-GlyRα1 Polyclonal Antibody (1:1000, AGR-001, Alomone Labs, Israel), Rabbit anti-gephyrin Polyclonal Antibody (1:2000, AIP-005, Alomone Labs, Israel), goat Anti-GlyRα3 polyclonal Antibody (1:500, SC-17282, Santa Cruz Biotechnology, Santa Cruz, CA, USA), or mouse anti-actin monoclonal antibody (1:10,000, MAB1501, Indianapolis, IN, USA) primary antibody to detect the expression of GlyRα1, GlyRα3 and gephyrin in the DRG and spinal cord.

Techniques: Western Blot, Injection, Expressing, Imaging

Journal: BMC Neuroscience

Article Title: Glycine receptors expression in rat spinal cord and dorsal root ganglion in prostaglandin E2 intrathecal injection models

doi: 10.1186/s12868-018-0470-8

Figure Lengend Snippet: a Western blot and quantitative analysis of b gephyrin, c GlyRα1, and d GlyRα3 protein expressions in L5 DRG at 1 h and 5 h after PGE2 intrathecal injection. The gephyrin, GlyRα1 and GlyRα3 protein expressions were significantly decreased in the PGE2 group at 5 h. Confocal microscopic imaging showed expression of e gephyrin, f GlyRα1 and g GlyRα3 localized mainly in the neuron cells. Each group had n = 3–6 rats. Scale Bars: 20 μm; ** p

Article Snippet: The membranes were firstly blocked with 5% milk in phosphate-buffered saline (PBS) with 0.1% Tween-20 for 1 h at room temperature, and then probed overnight at 4 °C with Rabbit anti-GlyRα1 Polyclonal Antibody (1:1000, AGR-001, Alomone Labs, Israel), Rabbit anti-gephyrin Polyclonal Antibody (1:2000, AIP-005, Alomone Labs, Israel), goat Anti-GlyRα3 polyclonal Antibody (1:500, SC-17282, Santa Cruz Biotechnology, Santa Cruz, CA, USA), or mouse anti-actin monoclonal antibody (1:10,000, MAB1501, Indianapolis, IN, USA) primary antibody to detect the expression of GlyRα1, GlyRα3 and gephyrin in the DRG and spinal cord.

Techniques: Western Blot, Injection, Imaging, Expressing

Journal: BMC Neuroscience

Article Title: Glycine receptors expression in rat spinal cord and dorsal root ganglion in prostaglandin E2 intrathecal injection models

doi: 10.1186/s12868-018-0470-8

Figure Lengend Snippet: Triple immunofluorescence staining showing GlyRα3, Gephyrin and NeuN co-localization in the L5 spinal cord. Positive for GlyRα3 are shown in green ( a ), positive for Gephyrin are shown in red ( b ) and positive for NeuN are shown in blue ( c ). Double-labelled images of GlyRα3 and Gephyrin ( d ), GlyRα3 and NeuN ( e ) are indicated. f The merged image demonstrates co-localization of GlyRα3, Gephyrin in Neurons. Scale bars = 200 µm, 20 µm

Article Snippet: The membranes were firstly blocked with 5% milk in phosphate-buffered saline (PBS) with 0.1% Tween-20 for 1 h at room temperature, and then probed overnight at 4 °C with Rabbit anti-GlyRα1 Polyclonal Antibody (1:1000, AGR-001, Alomone Labs, Israel), Rabbit anti-gephyrin Polyclonal Antibody (1:2000, AIP-005, Alomone Labs, Israel), goat Anti-GlyRα3 polyclonal Antibody (1:500, SC-17282, Santa Cruz Biotechnology, Santa Cruz, CA, USA), or mouse anti-actin monoclonal antibody (1:10,000, MAB1501, Indianapolis, IN, USA) primary antibody to detect the expression of GlyRα1, GlyRα3 and gephyrin in the DRG and spinal cord.

Techniques: Immunofluorescence, Staining

Journal: BMC Neuroscience

Article Title: Glycine receptors expression in rat spinal cord and dorsal root ganglion in prostaglandin E2 intrathecal injection models

doi: 10.1186/s12868-018-0470-8

Figure Lengend Snippet: Triple immunofluorescence staining showing GlyRα3, Gephyrin and NeuN co-localization in the L5 DRG. Positive for GlyRα3 are shown in green ( a ), positive for Gephyrin are shown in red ( b ) and positive for NeuN are shown in blue ( c ). Double-labelled images of GlyRα3 and Gephyrin ( d ), GlyRα3 and NeuN ( e ) are indicated. f The merged image demonstrates co-localization of GlyRα3, Gephyrin in Neurons of DRG. IF experiments from three different sham controls were performed. Secondary Abs only were included as negative controls. Scale bars = 20 µm

Article Snippet: The membranes were firstly blocked with 5% milk in phosphate-buffered saline (PBS) with 0.1% Tween-20 for 1 h at room temperature, and then probed overnight at 4 °C with Rabbit anti-GlyRα1 Polyclonal Antibody (1:1000, AGR-001, Alomone Labs, Israel), Rabbit anti-gephyrin Polyclonal Antibody (1:2000, AIP-005, Alomone Labs, Israel), goat Anti-GlyRα3 polyclonal Antibody (1:500, SC-17282, Santa Cruz Biotechnology, Santa Cruz, CA, USA), or mouse anti-actin monoclonal antibody (1:10,000, MAB1501, Indianapolis, IN, USA) primary antibody to detect the expression of GlyRα1, GlyRα3 and gephyrin in the DRG and spinal cord.

Techniques: Immunofluorescence, Staining