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    Cell Signaling Technology Inc nmda receptor 2b glun2b d15b3 rabbit mab
    Nmda Receptor 2b Glun2b D15b3 Rabbit Mab, supplied by Cell Signaling Technology Inc, 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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    95
    Alomone Labs rabbit rb antibodies against glun2b
    ECM digestion increases <t>p1472-GluN2B</t> level and decreases the endocytosis of GluN2B. ( A )Dissociated hippocampal cultures at DIV21-24 were treated with Hya over night and endocytosed GluN2B (green) was quantified using Map2 staining as mask (red). ( B ) There is less endocytosis of GluN2B after ECM removal within 30 minutes (Ctl 1.00 ± 0.02, n = 79; Hya 0.9 ± 0.02, n = 80; average ± SEM, Unpaired t-test, **P = 0.0015. Scale bar: 5 µm). ( C ) Quantitative WB from lysates of acute hippocampal slices treated with Ctl or Hya probed with an antibody against pGluN2B pTyr1472 (AP2 binding site) and GluN2B. ( D ) Quantification of WB of acute hippocampal slices and cortical cultures (DIV 21–24) revealed that the amount of phosphorylated GluN2B, normalized to the total amount of GluN2B, is increased after Hya treatment (overnight for cultures, 3 h for slices; slices: Ctl 1.00 ± 0.06, n = 4; Hya 1.23 ± 0.09, n = 4; cultures: Ctl 1.00 ± 0.05, n = 9; Hya 1.26 ± 0.1, n = 9; Unpaired t-test, cultures: P = 0.0332, slices P = 0.0837, ***P
    Rabbit Rb Antibodies Against Glun2b, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Alomone Labs glun2b blocking peptide
    Distribution of <t>GluN2B</t> immunoreactivity in young and aged GIN mice hippocampus. Panoramic confocal plane showing the distribution of O-LM cells (green) and GluN2B immunoreactivity (red) in the hippocampus of 3-month-old (A1) and 16-month-old (B1) mice. Different regions and strata are indicated with dotted lines. (A2,B2) High magnification view from the different CA1 strata in 3-month-old (A2) and 16-month-old (B2) mice. (A3,B3) Enlarged view of the squared regions in panels (A2,B2) , showing double immunofluorescence for GFP/GluN2B, in strata oriens , and pyramidale. Note the presence of GluN2B + clusters in pyramidal neurons in 16-month-old (B3) , but not in 3-month-old (A3) mice. Scale bar: 150 μm for panels (A1,B1 ), 67 μm for panels (A2,B2) , and 21 μm for panels (A3,B3) .
    Glun2b Blocking Peptide, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Cell Signaling Technology Inc n2b
    NMDAR mediated nNOS activation is prevented by LOPC at 15 days. ( A) Calcium accumulation in the thalamus is not affected by LOPC at all pixel intensities as shown by cumulative tresholding analysis (scalebar = 400 μm). ( B ) The thalamic expression NR1, <t>N2B</t> and nNOS do not change in time or with treatment as assessed by WB. At 15 days, WB analysis show that the expression of synaptic marker PSD-95 is decreased by 50% in LOPC while Synapsin 1 expression is unchanged. ( C ) Co-immunoprecipitation of N2B and PSD-95 shows decreased interaction in LOPC samples between NMDAR and PSD-95 at 15 days (-Ab: beads not coated incubated in lysate; -Lysate: beads coated incubated in PBS). Images cropped from Supp. Fig. 3 . Results are shown as the mean ± SD. *p
    N2b, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    ECM digestion increases p1472-GluN2B level and decreases the endocytosis of GluN2B. ( A )Dissociated hippocampal cultures at DIV21-24 were treated with Hya over night and endocytosed GluN2B (green) was quantified using Map2 staining as mask (red). ( B ) There is less endocytosis of GluN2B after ECM removal within 30 minutes (Ctl 1.00 ± 0.02, n = 79; Hya 0.9 ± 0.02, n = 80; average ± SEM, Unpaired t-test, **P = 0.0015. Scale bar: 5 µm). ( C ) Quantitative WB from lysates of acute hippocampal slices treated with Ctl or Hya probed with an antibody against pGluN2B pTyr1472 (AP2 binding site) and GluN2B. ( D ) Quantification of WB of acute hippocampal slices and cortical cultures (DIV 21–24) revealed that the amount of phosphorylated GluN2B, normalized to the total amount of GluN2B, is increased after Hya treatment (overnight for cultures, 3 h for slices; slices: Ctl 1.00 ± 0.06, n = 4; Hya 1.23 ± 0.09, n = 4; cultures: Ctl 1.00 ± 0.05, n = 9; Hya 1.26 ± 0.1, n = 9; Unpaired t-test, cultures: P = 0.0332, slices P = 0.0837, ***P

    Journal: Scientific Reports

    Article Title: Hyaluronic acid based extracellular matrix regulates surface expression of GluN2B containing NMDA receptors

    doi: 10.1038/s41598-017-07003-3

    Figure Lengend Snippet: ECM digestion increases p1472-GluN2B level and decreases the endocytosis of GluN2B. ( A )Dissociated hippocampal cultures at DIV21-24 were treated with Hya over night and endocytosed GluN2B (green) was quantified using Map2 staining as mask (red). ( B ) There is less endocytosis of GluN2B after ECM removal within 30 minutes (Ctl 1.00 ± 0.02, n = 79; Hya 0.9 ± 0.02, n = 80; average ± SEM, Unpaired t-test, **P = 0.0015. Scale bar: 5 µm). ( C ) Quantitative WB from lysates of acute hippocampal slices treated with Ctl or Hya probed with an antibody against pGluN2B pTyr1472 (AP2 binding site) and GluN2B. ( D ) Quantification of WB of acute hippocampal slices and cortical cultures (DIV 21–24) revealed that the amount of phosphorylated GluN2B, normalized to the total amount of GluN2B, is increased after Hya treatment (overnight for cultures, 3 h for slices; slices: Ctl 1.00 ± 0.06, n = 4; Hya 1.23 ± 0.09, n = 4; cultures: Ctl 1.00 ± 0.05, n = 9; Hya 1.26 ± 0.1, n = 9; Unpaired t-test, cultures: P = 0.0332, slices P = 0.0837, ***P

    Article Snippet: Antibodies and drugs The following commercial antibodies were used for Immunocytochemistry (ICC) and Western blot (WB) in the concentrations indicated: rabbit (rb) antibodies against GluN2B (alomone labs; ICC live staining: 1:200, fixed staining.

    Techniques: Staining, CTL Assay, Western Blot, Binding Assay

    ECM removal enhances GluN2B-NMDAR mediated synaptic currents. ( A ) Example traces of NMDAR - mediated sEPCSs before and after Hya treatment in dissociated hippocampal cultures DIV21-24. ( B ) Amplitudes of single peaks show no significant differences between Hya treated or Hya plus Ifenprodil treated cultures (Ctl, −905.5 ± 179.4, n = 10; Hya, −776.2 ± 174.8, n = 10; Hya + Ifen, −758.2 ± 161.7, n = 11; average ± SEM; One-way ANOVA, P = 0.7991). ( C ) Average of single peaks before and after Hya treatment and after Ifenprodil application. Normalization of the amplitude illustrates the increased decay-time after Hya treatment (red line) in comparison to Ctl (black line). This can be restored after Ifenprodil application (green line). Ctl traces are identical. ( D ) Quantification of the area under the curve (AUC) of averaged and normalized events (left), which represent the total charge transfer revealed bigger charge transfer after ECM removal, which was reduced to control levels after blocking GluN2B-NMDAR with Ifen (Ctl, 1 ± 0.02, n = 10; Hya, 1.38 ± 0.09, n = 10; Hya + Ifenprodil, 0.98 ± 0.05, n = 11; average ± SEM; One-way ANOVA, P

    Journal: Scientific Reports

    Article Title: Hyaluronic acid based extracellular matrix regulates surface expression of GluN2B containing NMDA receptors

    doi: 10.1038/s41598-017-07003-3

    Figure Lengend Snippet: ECM removal enhances GluN2B-NMDAR mediated synaptic currents. ( A ) Example traces of NMDAR - mediated sEPCSs before and after Hya treatment in dissociated hippocampal cultures DIV21-24. ( B ) Amplitudes of single peaks show no significant differences between Hya treated or Hya plus Ifenprodil treated cultures (Ctl, −905.5 ± 179.4, n = 10; Hya, −776.2 ± 174.8, n = 10; Hya + Ifen, −758.2 ± 161.7, n = 11; average ± SEM; One-way ANOVA, P = 0.7991). ( C ) Average of single peaks before and after Hya treatment and after Ifenprodil application. Normalization of the amplitude illustrates the increased decay-time after Hya treatment (red line) in comparison to Ctl (black line). This can be restored after Ifenprodil application (green line). Ctl traces are identical. ( D ) Quantification of the area under the curve (AUC) of averaged and normalized events (left), which represent the total charge transfer revealed bigger charge transfer after ECM removal, which was reduced to control levels after blocking GluN2B-NMDAR with Ifen (Ctl, 1 ± 0.02, n = 10; Hya, 1.38 ± 0.09, n = 10; Hya + Ifenprodil, 0.98 ± 0.05, n = 11; average ± SEM; One-way ANOVA, P

    Article Snippet: Antibodies and drugs The following commercial antibodies were used for Immunocytochemistry (ICC) and Western blot (WB) in the concentrations indicated: rabbit (rb) antibodies against GluN2B (alomone labs; ICC live staining: 1:200, fixed staining.

    Techniques: CTL Assay, Blocking Assay

    ECM removal leads to increased surface expression of GluN2B in a β1 - integrin dependent manner. ( A ) Dissociated hippocampal cultures were treated with Hya over night and stained against the total amount of GluN2B and the dendritic marker Map2 (scale bar: 10 μm. ( B ) Total GluN2B expression is not affected by ECM removal (Dendrites: Ctl 1 ± 0.10, n = 30; Hya 0.89 ± 0.03, n = 30, P = 0.31; Synapses: Ctl: 1 ± 0.03, n = 30; Hya: 1.05 ± 0.03, n = 30, P = 0.27; average ± SEM; unpaired t-test). ( C ) Quantitative WB of lysed cortical cultures (DIV21) pretreated with Hya over night show no significant change in GluN2B immunoreactivity. ( D ) Dissociated hippocampal cultures at DIV21-24 were treated with Hya over night and stained against surface GluN2B (green) and the synaptic marker PSD-95 (scale bar: 10 μm). ( E ) Synaptic GluN2B surface expression at various time points after Hya treatment (Ctl: 1 ± 0.04, n = 24; Hya 1,5 h: 1.08 ± 0.04, n = 22, P = 0.76; Hya 3 h: 1.40 ± 0.09, n = 30, P = 0.0001; Hya 6 h: 1.41 ± 0.13, n = 9, P = 0.002; Hya 12 h: 1.35 ± 0.08, n = 8, P = 0.01; Hya 48 h: 1.18 ± 0.05, n = 8, P = 0.04 average ± SEM; One way-ANOVA, Dunnett’s Multiple Comparison Test). ( F,G ) GluN2B surface expression at synapses and dendrites increases after ECM degradation and can be restored by simultaneous application of the β1-integrin function blocking antibody CD29. ( F ) Synapses: Ctl: 1.0 ± 0.05, n = 68; Hya: 1.3 ± 0.05, n = 70; Hya + CD29: 0.93 ± 0.03, n = 51. ( G ) Dendrites: Ctl 1.00 ± 0.04, n = 36; Hya 1.78 ± 0.11, n = 35; Hya + CD29 0.96 ± 0.03, n = 34; average ± SEM; One-way ANOVA, P

    Journal: Scientific Reports

    Article Title: Hyaluronic acid based extracellular matrix regulates surface expression of GluN2B containing NMDA receptors

    doi: 10.1038/s41598-017-07003-3

    Figure Lengend Snippet: ECM removal leads to increased surface expression of GluN2B in a β1 - integrin dependent manner. ( A ) Dissociated hippocampal cultures were treated with Hya over night and stained against the total amount of GluN2B and the dendritic marker Map2 (scale bar: 10 μm. ( B ) Total GluN2B expression is not affected by ECM removal (Dendrites: Ctl 1 ± 0.10, n = 30; Hya 0.89 ± 0.03, n = 30, P = 0.31; Synapses: Ctl: 1 ± 0.03, n = 30; Hya: 1.05 ± 0.03, n = 30, P = 0.27; average ± SEM; unpaired t-test). ( C ) Quantitative WB of lysed cortical cultures (DIV21) pretreated with Hya over night show no significant change in GluN2B immunoreactivity. ( D ) Dissociated hippocampal cultures at DIV21-24 were treated with Hya over night and stained against surface GluN2B (green) and the synaptic marker PSD-95 (scale bar: 10 μm). ( E ) Synaptic GluN2B surface expression at various time points after Hya treatment (Ctl: 1 ± 0.04, n = 24; Hya 1,5 h: 1.08 ± 0.04, n = 22, P = 0.76; Hya 3 h: 1.40 ± 0.09, n = 30, P = 0.0001; Hya 6 h: 1.41 ± 0.13, n = 9, P = 0.002; Hya 12 h: 1.35 ± 0.08, n = 8, P = 0.01; Hya 48 h: 1.18 ± 0.05, n = 8, P = 0.04 average ± SEM; One way-ANOVA, Dunnett’s Multiple Comparison Test). ( F,G ) GluN2B surface expression at synapses and dendrites increases after ECM degradation and can be restored by simultaneous application of the β1-integrin function blocking antibody CD29. ( F ) Synapses: Ctl: 1.0 ± 0.05, n = 68; Hya: 1.3 ± 0.05, n = 70; Hya + CD29: 0.93 ± 0.03, n = 51. ( G ) Dendrites: Ctl 1.00 ± 0.04, n = 36; Hya 1.78 ± 0.11, n = 35; Hya + CD29 0.96 ± 0.03, n = 34; average ± SEM; One-way ANOVA, P

    Article Snippet: Antibodies and drugs The following commercial antibodies were used for Immunocytochemistry (ICC) and Western blot (WB) in the concentrations indicated: rabbit (rb) antibodies against GluN2B (alomone labs; ICC live staining: 1:200, fixed staining.

    Techniques: Expressing, Staining, Marker, CTL Assay, Western Blot, Blocking Assay

    Distribution of GluN2B immunoreactivity in young and aged GIN mice hippocampus. Panoramic confocal plane showing the distribution of O-LM cells (green) and GluN2B immunoreactivity (red) in the hippocampus of 3-month-old (A1) and 16-month-old (B1) mice. Different regions and strata are indicated with dotted lines. (A2,B2) High magnification view from the different CA1 strata in 3-month-old (A2) and 16-month-old (B2) mice. (A3,B3) Enlarged view of the squared regions in panels (A2,B2) , showing double immunofluorescence for GFP/GluN2B, in strata oriens , and pyramidale. Note the presence of GluN2B + clusters in pyramidal neurons in 16-month-old (B3) , but not in 3-month-old (A3) mice. Scale bar: 150 μm for panels (A1,B1 ), 67 μm for panels (A2,B2) , and 21 μm for panels (A3,B3) .

    Journal: Frontiers in Aging Neuroscience

    Article Title: Effects of Aging on the Structure and Expression of NMDA Receptors of Somatostatin Expressing Neurons in the Mouse Hippocampus

    doi: 10.3389/fnagi.2021.782737

    Figure Lengend Snippet: Distribution of GluN2B immunoreactivity in young and aged GIN mice hippocampus. Panoramic confocal plane showing the distribution of O-LM cells (green) and GluN2B immunoreactivity (red) in the hippocampus of 3-month-old (A1) and 16-month-old (B1) mice. Different regions and strata are indicated with dotted lines. (A2,B2) High magnification view from the different CA1 strata in 3-month-old (A2) and 16-month-old (B2) mice. (A3,B3) Enlarged view of the squared regions in panels (A2,B2) , showing double immunofluorescence for GFP/GluN2B, in strata oriens , and pyramidale. Note the presence of GluN2B + clusters in pyramidal neurons in 16-month-old (B3) , but not in 3-month-old (A3) mice. Scale bar: 150 μm for panels (A1,B1 ), 67 μm for panels (A2,B2) , and 21 μm for panels (A3,B3) .

    Article Snippet: Controls were performed omitting the anti-GluN1 or anti-GluN2B antibody, as well as incubating with these antibodies previously pre-absorbed overnight with an excess of its immunogenic peptide (GluN1 blocking peptide, Alomone, Jerusalem, Israel) or (GluN2B blocking peptide, Alomone, Jerusalem, Israel), respectively.

    Techniques: Mouse Assay, Immunofluorescence

    Analysis of the density and percentage of area covered with GluN2B immunoreactive puncta in the somata and in the periphery of O-LM cells during aging. (A–F) Double GFP/GluN2B immunohistochemistry in 3-month-old (A) , 9-month-old (B) , 16-month-old (C) female mice and in 3-month-old (D) , 9-month-old (E) , 16-month-old (F) male mice. In panels (C2,F2) , a detailed view of the GluN2B clustering in aged mice can be observed. (G–I) Graphs showing the density and percentage of area covered with GluN2B immunoreactive puncta in the somata (G1,G2–I2) and in its periphery (G3,G4–I4) in animals segregated by sex (G1–4) , pooled females (H1–4) and males (I1–4) (all graphs represent mean ± SEM., * p -value

    Journal: Frontiers in Aging Neuroscience

    Article Title: Effects of Aging on the Structure and Expression of NMDA Receptors of Somatostatin Expressing Neurons in the Mouse Hippocampus

    doi: 10.3389/fnagi.2021.782737

    Figure Lengend Snippet: Analysis of the density and percentage of area covered with GluN2B immunoreactive puncta in the somata and in the periphery of O-LM cells during aging. (A–F) Double GFP/GluN2B immunohistochemistry in 3-month-old (A) , 9-month-old (B) , 16-month-old (C) female mice and in 3-month-old (D) , 9-month-old (E) , 16-month-old (F) male mice. In panels (C2,F2) , a detailed view of the GluN2B clustering in aged mice can be observed. (G–I) Graphs showing the density and percentage of area covered with GluN2B immunoreactive puncta in the somata (G1,G2–I2) and in its periphery (G3,G4–I4) in animals segregated by sex (G1–4) , pooled females (H1–4) and males (I1–4) (all graphs represent mean ± SEM., * p -value

    Article Snippet: Controls were performed omitting the anti-GluN1 or anti-GluN2B antibody, as well as incubating with these antibodies previously pre-absorbed overnight with an excess of its immunogenic peptide (GluN1 blocking peptide, Alomone, Jerusalem, Israel) or (GluN2B blocking peptide, Alomone, Jerusalem, Israel), respectively.

    Techniques: Immunohistochemistry, Mouse Assay

    NMDAR mediated nNOS activation is prevented by LOPC at 15 days. ( A) Calcium accumulation in the thalamus is not affected by LOPC at all pixel intensities as shown by cumulative tresholding analysis (scalebar = 400 μm). ( B ) The thalamic expression NR1, N2B and nNOS do not change in time or with treatment as assessed by WB. At 15 days, WB analysis show that the expression of synaptic marker PSD-95 is decreased by 50% in LOPC while Synapsin 1 expression is unchanged. ( C ) Co-immunoprecipitation of N2B and PSD-95 shows decreased interaction in LOPC samples between NMDAR and PSD-95 at 15 days (-Ab: beads not coated incubated in lysate; -Lysate: beads coated incubated in PBS). Images cropped from Supp. Fig. 3 . Results are shown as the mean ± SD. *p

    Journal: Scientific Reports

    Article Title: Low oxygen post conditioning prevents thalamic secondary neuronal loss caused by excitotoxicity after cortical stroke

    doi: 10.1038/s41598-019-39493-8

    Figure Lengend Snippet: NMDAR mediated nNOS activation is prevented by LOPC at 15 days. ( A) Calcium accumulation in the thalamus is not affected by LOPC at all pixel intensities as shown by cumulative tresholding analysis (scalebar = 400 μm). ( B ) The thalamic expression NR1, N2B and nNOS do not change in time or with treatment as assessed by WB. At 15 days, WB analysis show that the expression of synaptic marker PSD-95 is decreased by 50% in LOPC while Synapsin 1 expression is unchanged. ( C ) Co-immunoprecipitation of N2B and PSD-95 shows decreased interaction in LOPC samples between NMDAR and PSD-95 at 15 days (-Ab: beads not coated incubated in lysate; -Lysate: beads coated incubated in PBS). Images cropped from Supp. Fig. 3 . Results are shown as the mean ± SD. *p

    Article Snippet: After transfer and blocking, the membranes were probed with the appropriate antibody: CD11b (Cat#ab75476, Abcam), N2B (Cat#14544, Cell Signalling), NeuN (Cat#MAB377, Millipore), nNOS (Cat#4236, Cell Signalling), NR1 (Cat#5704, Cell Signalling), PSD-95 (Cat#3409, Cell Signalling), Synapsin 1 (Cat#5297, Cell Signalling), β-actin (Cat#A3854, Sigma-Aldrich).

    Techniques: Activation Assay, Expressing, Western Blot, Marker, Immunoprecipitation, Incubation