rabbit anti nr2a  (Millipore)


Bioz Verified Symbol Millipore is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 96
    Name:
    Anti Glutamate Receptor NMDAR2A NR2A antibody
    Description:
    N methyl D aspartate NMDA receptors are made up of NMDAR1 NR1 and NMDAR2A NMDAR2D NR2A NR2D subunits The glutamate receptor N methyl D aspartate 2A GRIN2A gene encodes the 2A subunit of the NMDA receptor The NMDA receptors are made up of NR1 and at least one subunit of the NR2 class forming a hetero pentamer complex
    Catalog Number:
    g9038
    Price:
    None
    Applications:
    Anti-Glutamate Receptor NMDAR2A (NR2A) antibody produced in rabbit is suitable as a primary antibody for immunoblotting using whole-tissue lysates or crude synaptosomal fractions of hippocampal slices from mice. It is suitable for immunoblotting at a working dilution of 1:3000 using a synaptosomal fraction of rat brain. It is also suitable for microarray analysis.
    Buy from Supplier


    Structured Review

    Millipore rabbit anti nr2a
    Expression of glutamate receptors. (A) Representative western-blot analysis of GluR1, <t>NR2A,</t> and NR2B subunits in the ACC homogenates. Levels of GluR1 (B) , NR2A (C), and NR2B (D) in saline control, vehicle-, T3-, and T4-treated HT mice. n = 5 in each group. * p
    N methyl D aspartate NMDA receptors are made up of NMDAR1 NR1 and NMDAR2A NMDAR2D NR2A NR2D subunits The glutamate receptor N methyl D aspartate 2A GRIN2A gene encodes the 2A subunit of the NMDA receptor The NMDA receptors are made up of NR1 and at least one subunit of the NR2 class forming a hetero pentamer complex
    https://www.bioz.com/result/rabbit anti nr2a/product/Millipore
    Average 96 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    rabbit anti nr2a - by Bioz Stars, 2020-08
    96/100 stars

    Images

    1) Product Images from "Decreased pain threshold and enhanced synaptic transmission in the anterior cingulate cortex of experimental hypothyroidism mice"

    Article Title: Decreased pain threshold and enhanced synaptic transmission in the anterior cingulate cortex of experimental hypothyroidism mice

    Journal: Molecular Pain

    doi: 10.1186/1744-8069-10-38

    Expression of glutamate receptors. (A) Representative western-blot analysis of GluR1, NR2A, and NR2B subunits in the ACC homogenates. Levels of GluR1 (B) , NR2A (C), and NR2B (D) in saline control, vehicle-, T3-, and T4-treated HT mice. n = 5 in each group. * p
    Figure Legend Snippet: Expression of glutamate receptors. (A) Representative western-blot analysis of GluR1, NR2A, and NR2B subunits in the ACC homogenates. Levels of GluR1 (B) , NR2A (C), and NR2B (D) in saline control, vehicle-, T3-, and T4-treated HT mice. n = 5 in each group. * p

    Techniques Used: Expressing, Western Blot, Mouse Assay

    2) Product Images from "IQGAP1 regulates NR2A signaling, spine density, and cognitive processes"

    Article Title: IQGAP1 regulates NR2A signaling, spine density, and cognitive processes

    Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

    doi: 10.1523/JNEUROSCI.1300-11.2011

    IQGAP1 interacts with NR2 subunits and regulates their surface expression. A , IQGAP1 interacted with NR2A, and to a lesser extent NR2B (top) but not NR3A or NR3B (bottom). B , IQGAP1 also interacted with PSD-95. C–E , Representative immunoblots
    Figure Legend Snippet: IQGAP1 interacts with NR2 subunits and regulates their surface expression. A , IQGAP1 interacted with NR2A, and to a lesser extent NR2B (top) but not NR3A or NR3B (bottom). B , IQGAP1 also interacted with PSD-95. C–E , Representative immunoblots

    Techniques Used: Expressing, Western Blot

    3) Product Images from "Reelin Exerts Structural, Biochemical and Transcriptional Regulation Over Presynaptic and Postsynaptic Elements in the Adult Hippocampus"

    Article Title: Reelin Exerts Structural, Biochemical and Transcriptional Regulation Over Presynaptic and Postsynaptic Elements in the Adult Hippocampus

    Journal: Frontiers in Cellular Neuroscience

    doi: 10.3389/fncel.2016.00138

    Expression of NMDA receptor subunits NR2a and NR2b in Reelin-OE mice. (A) Left, immunoblot analysis of NR2a in control, Reelin-OE, and DOX-treated Reelin-OE mice. Right, histogram showing densitometric analysis ( n = 4 animals per group) by fold change in the three groups of mice (mean ± SEM; ANOVA test). (B) Left, immunoblot analysis of NR2b in control, Reelin-OE, and DOX-treated Reelin-OE mice. Right, histogram showing densitometric analysis ( n = 3 animals per group) by fold change in the three groups of mice (mean ± SEM; ANOVA test). (C–E) Examples of immunogold labeling against NR2a in dendritic spines (s) receiving a synaptic contact from an axon terminal (at) in the SR of control (C) , Reelin-OE (D) , and DOX-treated Reelin-OE (E) mice. (F) Histogram illustrating the proportion of NR2a-gold particles in the synaptic contact, the extrasynaptic membrane and the intracellular domain of dendritic spines in the SR of Reelin-OE and DOX-treated Reelin-OE mice and their control littermates (mean ± SEM; * p
    Figure Legend Snippet: Expression of NMDA receptor subunits NR2a and NR2b in Reelin-OE mice. (A) Left, immunoblot analysis of NR2a in control, Reelin-OE, and DOX-treated Reelin-OE mice. Right, histogram showing densitometric analysis ( n = 4 animals per group) by fold change in the three groups of mice (mean ± SEM; ANOVA test). (B) Left, immunoblot analysis of NR2b in control, Reelin-OE, and DOX-treated Reelin-OE mice. Right, histogram showing densitometric analysis ( n = 3 animals per group) by fold change in the three groups of mice (mean ± SEM; ANOVA test). (C–E) Examples of immunogold labeling against NR2a in dendritic spines (s) receiving a synaptic contact from an axon terminal (at) in the SR of control (C) , Reelin-OE (D) , and DOX-treated Reelin-OE (E) mice. (F) Histogram illustrating the proportion of NR2a-gold particles in the synaptic contact, the extrasynaptic membrane and the intracellular domain of dendritic spines in the SR of Reelin-OE and DOX-treated Reelin-OE mice and their control littermates (mean ± SEM; * p

    Techniques Used: Expressing, Mouse Assay, Labeling

    4) Product Images from "Odorant Deprivation Reversibly Modulates Transsynaptic Changes in the NR2B-Mediated CREB Pathway in Mouse Piriform Cortex"

    Article Title: Odorant Deprivation Reversibly Modulates Transsynaptic Changes in the NR2B-Mediated CREB Pathway in Mouse Piriform Cortex

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.1727-06.2006

    Expression of glutamate receptors in OB and PC after bilateral zinc sulfate lesion. The amount of NR2B and NR2A mRNA expression was measured by semiquantitative RT-PCR. A , B , NR2B expression declined 7 d after bilateral zinc sulfate lesion (BZL) ( A ) in OB, aPC, and pPC [two way ANOVA across all tissues and lesions, F (4, 30) = 0.91, p = 0.47, no interaction between lesion and tissue; one-way ANOVA for the main effect of the lesion, F (1, 30) = 25.18, p
    Figure Legend Snippet: Expression of glutamate receptors in OB and PC after bilateral zinc sulfate lesion. The amount of NR2B and NR2A mRNA expression was measured by semiquantitative RT-PCR. A , B , NR2B expression declined 7 d after bilateral zinc sulfate lesion (BZL) ( A ) in OB, aPC, and pPC [two way ANOVA across all tissues and lesions, F (4, 30) = 0.91, p = 0.47, no interaction between lesion and tissue; one-way ANOVA for the main effect of the lesion, F (1, 30) = 25.18, p

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction

    5) Product Images from "Neurological effects of inorganic arsenic exposure: altered cysteine/glutamate transport, NMDA expression and spatial memory impairment"

    Article Title: Neurological effects of inorganic arsenic exposure: altered cysteine/glutamate transport, NMDA expression and spatial memory impairment

    Journal: Frontiers in Cellular Neuroscience

    doi: 10.3389/fncel.2015.00021

    Cystine, cysteine and glutamate transporters in male hippocampus on PND 90 . (A) xCT; (B) EAAC1; (C) GLT1; (D) GLAST; (E) NR2A; (F) NR2B. Densitometric evaluation of the blot images was performed using β-tubulin as loading control. Bars represent mean ± SE relative to control values. Data were analyzed using one-way ANOVA and Tukey’s post hoc test. (*) Significantly different from controls, P
    Figure Legend Snippet: Cystine, cysteine and glutamate transporters in male hippocampus on PND 90 . (A) xCT; (B) EAAC1; (C) GLT1; (D) GLAST; (E) NR2A; (F) NR2B. Densitometric evaluation of the blot images was performed using β-tubulin as loading control. Bars represent mean ± SE relative to control values. Data were analyzed using one-way ANOVA and Tukey’s post hoc test. (*) Significantly different from controls, P

    Techniques Used:

    Cystine/cysteine and glutamate transporters expression in control and gestational exposed mice on PND 15 . Confocal images of (A) xCT and (B) EAAC1 expression in control and PND 15 CA1 hippocampal cells. Neuron marker anti-MAP2 (green), xCT (red) or EAAC1 (red). Nucleus were counterstained with DAPI (blue). Scale bar, 30 μm. The cortex and hippocampus regions were removed from males and females and processed for western blotting as described in Materials and Methods for (C) xCT; (D) EAAC1; (E) NMDAR NR2A and; (F) NMDAR NR2B subunits. Densitometric evaluation of the blot images was performed using β-tubulin as loading control. Bars represent mean ± SE relative to control values, n = 4–6. Data were analyzed using Student’s t -test. (*) Significantly different from controls,* P
    Figure Legend Snippet: Cystine/cysteine and glutamate transporters expression in control and gestational exposed mice on PND 15 . Confocal images of (A) xCT and (B) EAAC1 expression in control and PND 15 CA1 hippocampal cells. Neuron marker anti-MAP2 (green), xCT (red) or EAAC1 (red). Nucleus were counterstained with DAPI (blue). Scale bar, 30 μm. The cortex and hippocampus regions were removed from males and females and processed for western blotting as described in Materials and Methods for (C) xCT; (D) EAAC1; (E) NMDAR NR2A and; (F) NMDAR NR2B subunits. Densitometric evaluation of the blot images was performed using β-tubulin as loading control. Bars represent mean ± SE relative to control values, n = 4–6. Data were analyzed using Student’s t -test. (*) Significantly different from controls,* P

    Techniques Used: Expressing, Mouse Assay, Marker, Western Blot

    6) Product Images from "In vivo composition of NMDA receptor signalling complexes differs between membrane subdomains and is modulated by PSD-95 and PSD-93"

    Article Title: In vivo composition of NMDA receptor signalling complexes differs between membrane subdomains and is modulated by PSD-95 and PSD-93

    Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

    doi: 10.1523/JNEUROSCI.1792-10.2010

    Analysis of mice carrying a C-terminal truncation of NR2A subunits ( NR2A ΔC/ΔC ) Cortices of wild type and NR2A ΔC/ΔC litter-matched mice were dissected and lipid rafts were isolated by density gradient. A) Equal amounts of proteins from lipid raft fractions were analyzed by western blot for indicated proteins and equal volumes were analyzed for GM1 (dot blot) and cholesterol concentration (spectrophotometric determination). B) Equal amounts of protein from the PSD fraction were analyzed by western blot for the indicated proteins. C) Total proteins in crude homogenate were analyzed by western blot. D) Total proteins in the detergent-soluble extract were analyzed by western-blot. Graphics show densitometry analysis of blots (mean ± SEM) expressed as the ratio of immunoreactivity ( NR2A ΔC/ΔC mice over wild types) for six independent pairs of litter-matched mice, except (D) detergent-soluble extract (4 litter matched pairs of animals), ** p
    Figure Legend Snippet: Analysis of mice carrying a C-terminal truncation of NR2A subunits ( NR2A ΔC/ΔC ) Cortices of wild type and NR2A ΔC/ΔC litter-matched mice were dissected and lipid rafts were isolated by density gradient. A) Equal amounts of proteins from lipid raft fractions were analyzed by western blot for indicated proteins and equal volumes were analyzed for GM1 (dot blot) and cholesterol concentration (spectrophotometric determination). B) Equal amounts of protein from the PSD fraction were analyzed by western blot for the indicated proteins. C) Total proteins in crude homogenate were analyzed by western blot. D) Total proteins in the detergent-soluble extract were analyzed by western-blot. Graphics show densitometry analysis of blots (mean ± SEM) expressed as the ratio of immunoreactivity ( NR2A ΔC/ΔC mice over wild types) for six independent pairs of litter-matched mice, except (D) detergent-soluble extract (4 litter matched pairs of animals), ** p

    Techniques Used: Mouse Assay, Isolation, Western Blot, Dot Blot, Concentration Assay

    7) Product Images from "Genetic Deletion of NR3A Accelerates Glutamatergic Synapse Maturation"

    Article Title: Genetic Deletion of NR3A Accelerates Glutamatergic Synapse Maturation

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0042327

    Glutamate receptor subunits NR1, NR2A, NR2B, and GluR1 are highly enriched in PSDs of postnatal mice. Relative protein levels of ( A ) NR1, ( B ) NR2A, ( C ) NR2B, and ( D ) GluR1 in mouse forebrain. Protein data are averaged means of immunoreactive (IR) values relative to total protein loads (µg). n = 5–9. Error bars represent SEM. * p
    Figure Legend Snippet: Glutamate receptor subunits NR1, NR2A, NR2B, and GluR1 are highly enriched in PSDs of postnatal mice. Relative protein levels of ( A ) NR1, ( B ) NR2A, ( C ) NR2B, and ( D ) GluR1 in mouse forebrain. Protein data are averaged means of immunoreactive (IR) values relative to total protein loads (µg). n = 5–9. Error bars represent SEM. * p

    Techniques Used: Mouse Assay

    Deletion of NR3A transiently accelerates expression of synapse maturation markers. ( A ) Representative immunoblots from NR3A-KO compared to WT controls show increased PSD levels of ( B ) NR1, ( C ) NR2A, and ( D ) GluR1 at P8 that return to WT levels by P16 and adult ages. ( E ) NR2B expression is unchanged in the NR3A-KO. ( B, C, D, E ) Data are averaged means of immunoreactive values relative to protein loads (µg) and presented as percent of control values. NR1 values for age P8 are re-plotted here from [14] for comparative purposes. Error bars represent SEM. n = 7–10. Significance from control: * p
    Figure Legend Snippet: Deletion of NR3A transiently accelerates expression of synapse maturation markers. ( A ) Representative immunoblots from NR3A-KO compared to WT controls show increased PSD levels of ( B ) NR1, ( C ) NR2A, and ( D ) GluR1 at P8 that return to WT levels by P16 and adult ages. ( E ) NR2B expression is unchanged in the NR3A-KO. ( B, C, D, E ) Data are averaged means of immunoreactive values relative to protein loads (µg) and presented as percent of control values. NR1 values for age P8 are re-plotted here from [14] for comparative purposes. Error bars represent SEM. n = 7–10. Significance from control: * p

    Techniques Used: Expressing, Western Blot

    8) Product Images from "ApoE isoform-dependent changes in hippocampal synaptic function"

    Article Title: ApoE isoform-dependent changes in hippocampal synaptic function

    Journal: Molecular Neurodegeneration

    doi: 10.1186/1750-1326-4-21

    Effect of acute apoE exposure on NR2A and NR2B tyrosine phosphorylation . ApoE-deficient slices treated with 100 nM rhapoE isoforms. A) Ratio of pNR2A as measured by pTyr immunoreactivity to NR2A immunoreactivity as immunoprecipatated by NR2A. B) Ratio of pNR2B as measured by pTyr immunoreactivity to NR2B immunoreactivity as immunoprecipatated by NR2B. control (C, light grey), rhapoE2 (E2, medium grey), rhapoE3 (E3, dark grey), rhapoE4 (E4, black). Data expressed as mean ± SEM.
    Figure Legend Snippet: Effect of acute apoE exposure on NR2A and NR2B tyrosine phosphorylation . ApoE-deficient slices treated with 100 nM rhapoE isoforms. A) Ratio of pNR2A as measured by pTyr immunoreactivity to NR2A immunoreactivity as immunoprecipatated by NR2A. B) Ratio of pNR2B as measured by pTyr immunoreactivity to NR2B immunoreactivity as immunoprecipatated by NR2B. control (C, light grey), rhapoE2 (E2, medium grey), rhapoE3 (E3, dark grey), rhapoE4 (E4, black). Data expressed as mean ± SEM.

    Techniques Used:

    Effect of chronic apoE isoform expression on NR2A, NR2B levels and tyrosine phosphorylation . A) Levels of NR2A immunoreactivity, standardized to NR1 immunoreactivity, from CA1 of apoE TR animals. B) Levels of NR2B immunoreactivity, standardized to NR1 immunoreactivity, from CA1 of apoE TR animals. C) Levels of phosphotyrosine immunoreactivity at the molecular weight corresponding to NR2A and NR2B from CA1 of apoE TR animals. apoE2 TR (E2, medium grey), apoE3 TR (E3, dark grey), apoE4 TR (E4, black). Data expressed as mean ± SEM.
    Figure Legend Snippet: Effect of chronic apoE isoform expression on NR2A, NR2B levels and tyrosine phosphorylation . A) Levels of NR2A immunoreactivity, standardized to NR1 immunoreactivity, from CA1 of apoE TR animals. B) Levels of NR2B immunoreactivity, standardized to NR1 immunoreactivity, from CA1 of apoE TR animals. C) Levels of phosphotyrosine immunoreactivity at the molecular weight corresponding to NR2A and NR2B from CA1 of apoE TR animals. apoE2 TR (E2, medium grey), apoE3 TR (E3, dark grey), apoE4 TR (E4, black). Data expressed as mean ± SEM.

    Techniques Used: Expressing, Molecular Weight

    9) Product Images from "PSD-93 Deficiency Protects Cultured Cortical Neurons from NMDA Receptor-triggered Neurotoxicity"

    Article Title: PSD-93 Deficiency Protects Cultured Cortical Neurons from NMDA Receptor-triggered Neurotoxicity

    Journal: Neuroscience

    doi: 10.1016/j.neuroscience.2010.01.030

    PSD-93 deficiency significantly reduces synaptic expression of NR2A and NR2B in cultured cortical neurons. (A) Representative Western blots showing the levels of PSD-93, PSD-95, NR2A, and NR2B in the total soluble and synaptosomal fractions. (B) Statistical summary of the densitometric analysis expressed relative to WT mice after normalization to corresponding β-actin or N-cadherin. ** P
    Figure Legend Snippet: PSD-93 deficiency significantly reduces synaptic expression of NR2A and NR2B in cultured cortical neurons. (A) Representative Western blots showing the levels of PSD-93, PSD-95, NR2A, and NR2B in the total soluble and synaptosomal fractions. (B) Statistical summary of the densitometric analysis expressed relative to WT mice after normalization to corresponding β-actin or N-cadherin. ** P

    Techniques Used: Expressing, Cell Culture, Western Blot, Mouse Assay

    10) Product Images from "Inhibition of Soluble Tumor Necrosis Factor Ameliorates Synaptic Alterations and Ca2+ Dysregulation in Aged Rats"

    Article Title: Inhibition of Soluble Tumor Necrosis Factor Ameliorates Synaptic Alterations and Ca2+ Dysregulation in Aged Rats

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0038170

    Effects of XPro1595 on glutamate receptor protein levels. A, Representative Western blots are shown for AMPA receptor (GluR1 and GluR2) and NMDA receptor subtypes (NR1, NR2A, and NR2B) in hippocampal membrane fractions from aged (22 month) rats treated for four weeks (intraventricular delivery) with vehicle or XPro1595 (0.08 mg/kg/day). The Na/K-ATPase loading control is shown below. B, XPro1595 treatment resulted in a selective increase in GluR1 levels (* p
    Figure Legend Snippet: Effects of XPro1595 on glutamate receptor protein levels. A, Representative Western blots are shown for AMPA receptor (GluR1 and GluR2) and NMDA receptor subtypes (NR1, NR2A, and NR2B) in hippocampal membrane fractions from aged (22 month) rats treated for four weeks (intraventricular delivery) with vehicle or XPro1595 (0.08 mg/kg/day). The Na/K-ATPase loading control is shown below. B, XPro1595 treatment resulted in a selective increase in GluR1 levels (* p

    Techniques Used: Western Blot

    11) Product Images from "Anxiolytic-like effects of translocator protein (TSPO) ligand ZBD-2 in an animal model of chronic pain"

    Article Title: Anxiolytic-like effects of translocator protein (TSPO) ligand ZBD-2 in an animal model of chronic pain

    Journal: Molecular Pain

    doi: 10.1186/s12990-015-0013-6

    Effects of ZBD-2 on protein expression in the BLA. (A) Representative results of Western blot analysis in the BLA on the Day 21 after hindpaw CFA-injection. (B) ZBD-2 (0.15 and 1.5 mg/kg) for one week slightly decreased the levels of NR2A-containing NMDARs. (C) ZBD-2 (1.5 mg/kg) reversed the upregulation of NR2B-containing NMDARs. (D-E) ZBD-2 (0.15 and 1.5 mg/kg) reversed the upregulation of GluR1 and CaMKII-α. (F) Representative results of Western blot analysis for GABA A -α2 and GAD67. (G and H) Levels of GABA A -α2 and GAD67 were not altered in groups. n = 6 in each group, * p
    Figure Legend Snippet: Effects of ZBD-2 on protein expression in the BLA. (A) Representative results of Western blot analysis in the BLA on the Day 21 after hindpaw CFA-injection. (B) ZBD-2 (0.15 and 1.5 mg/kg) for one week slightly decreased the levels of NR2A-containing NMDARs. (C) ZBD-2 (1.5 mg/kg) reversed the upregulation of NR2B-containing NMDARs. (D-E) ZBD-2 (0.15 and 1.5 mg/kg) reversed the upregulation of GluR1 and CaMKII-α. (F) Representative results of Western blot analysis for GABA A -α2 and GAD67. (G and H) Levels of GABA A -α2 and GAD67 were not altered in groups. n = 6 in each group, * p

    Techniques Used: Expressing, Western Blot, Injection

    12) Product Images from "Development and characterization of NEX-Pten, a novel forebrain excitatory neuron-specific knockout mouse"

    Article Title: Development and characterization of NEX-Pten, a novel forebrain excitatory neuron-specific knockout mouse

    Journal: Developmental neuroscience

    doi: 10.1159/000337229

    Abnormal expression of proteins involved in brain development and synaptic plasticity in the forebrain of newborn homozygous NEX- Pten mice. Representative Western blots show upregulation of Dab1 ( a ), MAP2 and NeuN ( c ), NR2A, NR2B and GluR1 ( e ) in homozygous (Hom) compared to heterozygous (Het) and wild type (WT) mice. Quantitative analysis of data obtained from 5–6 mice per genotype confirms that Dab1 ( b ), MAP2 ( d ), NR2A and NR2B levels ( f ) normalized to actin are significantly higher in homozygous than in wild type samples ( p
    Figure Legend Snippet: Abnormal expression of proteins involved in brain development and synaptic plasticity in the forebrain of newborn homozygous NEX- Pten mice. Representative Western blots show upregulation of Dab1 ( a ), MAP2 and NeuN ( c ), NR2A, NR2B and GluR1 ( e ) in homozygous (Hom) compared to heterozygous (Het) and wild type (WT) mice. Quantitative analysis of data obtained from 5–6 mice per genotype confirms that Dab1 ( b ), MAP2 ( d ), NR2A and NR2B levels ( f ) normalized to actin are significantly higher in homozygous than in wild type samples ( p

    Techniques Used: Expressing, Mouse Assay, Western Blot

    13) Product Images from "Neuregulin and BDNF Induce a Switch to NMDA Receptor-Dependent Myelination by Oligodendrocytes"

    Article Title: Neuregulin and BDNF Induce a Switch to NMDA Receptor-Dependent Myelination by Oligodendrocytes

    Journal: PLoS Biology

    doi: 10.1371/journal.pbio.1001743

    Expression of NMDA receptors. (A) Western blots of control and NRG-treated cocultures for NR1, NR2B and its phosphorylated form (pNR2B), NR2C and its phosphorylated form (pNR2C), NR3A and NR3B, as well as for NR2A and NR2D compared to their respective positive controls of rat cortex (Ctx) and thalamus (Th); β-actin acts as a loading control throughout. (B) Densitometric quantification of subunit protein levels in cocultures (normalized to β-actin) in NRG normalized to the levels in control (NR2A and NR2D levels were undetectable). (C) Western blot of control (Con) and NRG-treated pure DRG cultures for NR3A and (below) densitometric quantification of subunit protein levels (normalized to β-actin and then to control). (D) Western blot for NR1 and NR3A of control (Con) and NRG-treated (for 6 d) pure OPC cultures, treated (+) or not treated (−) with 20 min glutamate (Glu, 100 µM) stimulation every day, with densitometric quantification of subunit protein levels (normalized to β-actin and then to control). The p values over the bars, in (B) from Holm–Bonferroni corrected t tests and in (C and D) from one-sample Student t tests, compare with control; numbers of experiments shown on bars.
    Figure Legend Snippet: Expression of NMDA receptors. (A) Western blots of control and NRG-treated cocultures for NR1, NR2B and its phosphorylated form (pNR2B), NR2C and its phosphorylated form (pNR2C), NR3A and NR3B, as well as for NR2A and NR2D compared to their respective positive controls of rat cortex (Ctx) and thalamus (Th); β-actin acts as a loading control throughout. (B) Densitometric quantification of subunit protein levels in cocultures (normalized to β-actin) in NRG normalized to the levels in control (NR2A and NR2D levels were undetectable). (C) Western blot of control (Con) and NRG-treated pure DRG cultures for NR3A and (below) densitometric quantification of subunit protein levels (normalized to β-actin and then to control). (D) Western blot for NR1 and NR3A of control (Con) and NRG-treated (for 6 d) pure OPC cultures, treated (+) or not treated (−) with 20 min glutamate (Glu, 100 µM) stimulation every day, with densitometric quantification of subunit protein levels (normalized to β-actin and then to control). The p values over the bars, in (B) from Holm–Bonferroni corrected t tests and in (C and D) from one-sample Student t tests, compare with control; numbers of experiments shown on bars.

    Techniques Used: Expressing, Western Blot

    14) Product Images from "NMDA NR2A and NR2B Receptors in the Rostral Anterior Cingulate Cortex Contribute to Pain-related Aversion in Male Rats"

    Article Title: NMDA NR2A and NR2B Receptors in the Rostral Anterior Cingulate Cortex Contribute to Pain-related Aversion in Male Rats

    Journal: Pain

    doi: 10.1016/j.pain.2009.07.027

    Effects of blocking NMDA receptor NR2A or NR2B subunits on formalin nociceptive conditioning-induced Fos expression in the rACC A, Schematic of experimental designed in subsequent studies. B, Photomicrographs showing expression of Fos on the rACC at 2 hrs after i.pl. injection of NS and formalin. C, Schematic of experimental designed in subsequent studies. D, Western blot for Fos from the rACC representative of 2 hrs after i.pl. injection of NS and formalin with contextual conditioning exposure. E, Densitometry analysis showing a significant increase in the Fos expression in the rACC following i.pl. injection of formalin with contextual conditioning exposure, which was significantly attenuated by intra-rACC injection of NR2B antagonists or NR2A AS ODNs. Two sides of the rACC were pooled in this experiment. Tubulin served as loading control. *p
    Figure Legend Snippet: Effects of blocking NMDA receptor NR2A or NR2B subunits on formalin nociceptive conditioning-induced Fos expression in the rACC A, Schematic of experimental designed in subsequent studies. B, Photomicrographs showing expression of Fos on the rACC at 2 hrs after i.pl. injection of NS and formalin. C, Schematic of experimental designed in subsequent studies. D, Western blot for Fos from the rACC representative of 2 hrs after i.pl. injection of NS and formalin with contextual conditioning exposure. E, Densitometry analysis showing a significant increase in the Fos expression in the rACC following i.pl. injection of formalin with contextual conditioning exposure, which was significantly attenuated by intra-rACC injection of NR2B antagonists or NR2A AS ODNs. Two sides of the rACC were pooled in this experiment. Tubulin served as loading control. *p

    Techniques Used: Blocking Assay, Expressing, Injection, Western Blot

    Immunohistochemistry for NMDA receptor NR1, NR2A, and NR2B subunits on the rACC A, Photomicrographs showing expression of NR1 on the contralateral rACC at 6 hrs after i.pl. injection of NS and formalin. Low, double immunofluorescence showing that NR1 (green) is co-localized with NeuN (neuronal marker, red), but not with GFAP (astrocytic marker, red), and OX-42 (microglial marker, red) in the rACC. B, Quantification of NR1-positive cells on both sides of the rACC. C,E, Photomicrographs showing expression of NR2A (C) and NR2B (E) on the contralateral rACC at 6 hrs after i.pl. injection of NS and formalin. Right, double immunofluorescence showing that either NR2A (C, green) or NR2B (E, green) is co-localized with NeuN (red) in the rACC. D,F, Quantification of NR2A-(D) and NR2B-(F) IP cells on both sides of the rACC. *p
    Figure Legend Snippet: Immunohistochemistry for NMDA receptor NR1, NR2A, and NR2B subunits on the rACC A, Photomicrographs showing expression of NR1 on the contralateral rACC at 6 hrs after i.pl. injection of NS and formalin. Low, double immunofluorescence showing that NR1 (green) is co-localized with NeuN (neuronal marker, red), but not with GFAP (astrocytic marker, red), and OX-42 (microglial marker, red) in the rACC. B, Quantification of NR1-positive cells on both sides of the rACC. C,E, Photomicrographs showing expression of NR2A (C) and NR2B (E) on the contralateral rACC at 6 hrs after i.pl. injection of NS and formalin. Right, double immunofluorescence showing that either NR2A (C, green) or NR2B (E, green) is co-localized with NeuN (red) in the rACC. D,F, Quantification of NR2A-(D) and NR2B-(F) IP cells on both sides of the rACC. *p

    Techniques Used: Immunohistochemistry, Expressing, Injection, Immunofluorescence, Marker

    Western blot for NMDA receptor NR1, NR2A, and NR2B subunits from the rACC representative of 6 hrs after i.pl. injection of NS and formalin with or without contextual conditioning exposure A, Schematic of experimental designed in subsequent studies. B–G Western blot analysis revealing a significant increase in NR2A (D,E) and NR2B (F,G), but not NR1 (B,C) in the rACC following i.pl. formalin injection with or without contextual conditioning exposure. Two sides of the rACC were pooled in this experiment. Tubulin served as loading control. *p
    Figure Legend Snippet: Western blot for NMDA receptor NR1, NR2A, and NR2B subunits from the rACC representative of 6 hrs after i.pl. injection of NS and formalin with or without contextual conditioning exposure A, Schematic of experimental designed in subsequent studies. B–G Western blot analysis revealing a significant increase in NR2A (D,E) and NR2B (F,G), but not NR1 (B,C) in the rACC following i.pl. formalin injection with or without contextual conditioning exposure. Two sides of the rACC were pooled in this experiment. Tubulin served as loading control. *p

    Techniques Used: Western Blot, Injection

    NMDA receptor NR2A and NR2B subunits contribute to the induction of F-CPA A, Schematic of the protocol for behavioral testing. B,C, Histograms showing formalin-induced CPA. Mean ± SEM time spent in the treatment-paired compartment on pre-and post-conditioning day (B). Mean ± SEM F-CPA scores, i.e. the time spent in the treatment-paired compartment on the pre-conditioning day minus that on the post-conditioning day (C). D, Schematic of the behavioral procedure used in subsequent studies. E,F, Histograms showing effects of microinjection of NR2B antagonist Ifenprodil (0.2 μg/μl, 0.6 μl per side), Ro 25-6981 (2 μg/μl, 0.6 μl per side), or NR2A antisense (AS) ODNs (2 nmol/0.6 μl per side) into the bilateral rACC on F-CPA. Values are mean ± SEM time in the formalin-paired compartment on pre-conditioning and post-conditioning days (E), and F-CPA scores (F). G,H, Western blot analysis showing that formalin-induced increase in level of NR2A receptor is blocked by intra-rACC preadministration of NR2A AS but not NR2A MS ODNs. Tubulin serves as loading control. Both ipsilateral and contralateral sides of the rACC were pooled together in this experiment. I,J, Intra-rACC administration of NR2B antagonist Ifenprodil and NS (I), or NR2A AS and missense (MS) ODNs (J) has no effect on formalin-induced biphasic nociceptive responses. *p
    Figure Legend Snippet: NMDA receptor NR2A and NR2B subunits contribute to the induction of F-CPA A, Schematic of the protocol for behavioral testing. B,C, Histograms showing formalin-induced CPA. Mean ± SEM time spent in the treatment-paired compartment on pre-and post-conditioning day (B). Mean ± SEM F-CPA scores, i.e. the time spent in the treatment-paired compartment on the pre-conditioning day minus that on the post-conditioning day (C). D, Schematic of the behavioral procedure used in subsequent studies. E,F, Histograms showing effects of microinjection of NR2B antagonist Ifenprodil (0.2 μg/μl, 0.6 μl per side), Ro 25-6981 (2 μg/μl, 0.6 μl per side), or NR2A antisense (AS) ODNs (2 nmol/0.6 μl per side) into the bilateral rACC on F-CPA. Values are mean ± SEM time in the formalin-paired compartment on pre-conditioning and post-conditioning days (E), and F-CPA scores (F). G,H, Western blot analysis showing that formalin-induced increase in level of NR2A receptor is blocked by intra-rACC preadministration of NR2A AS but not NR2A MS ODNs. Tubulin serves as loading control. Both ipsilateral and contralateral sides of the rACC were pooled together in this experiment. I,J, Intra-rACC administration of NR2B antagonist Ifenprodil and NS (I), or NR2A AS and missense (MS) ODNs (J) has no effect on formalin-induced biphasic nociceptive responses. *p

    Techniques Used: Western Blot, Mass Spectrometry

    NMDA receptor NR2A and NR2B subunits fail to affect the induction of S-CPA A, Schematic of the protocol for behavioral testing. B,C, Histograms showing low-intensity (0.5 mA, 2 s) electric foot shock-induced CPA. Mean ± SEM time spent in the treatment-paired compartment on pre- and post-conditioning day (B). Mean ± SEM S-CPA scores, i.e. the time spent in the treatment-paired compartment on the pre-conditioning day minus that on the post-conditioning day (C). D, Schematic of the behavioral procedure used in subsequent studies. E,F, Histograms showing effects of microinjection of NR2B antagonist Ifenprodil (0.2 μg/μl, 0.6 μl per side), Ro 25-6981 (2 μg/μl, 0.6 μl per side), or NR2A AS ODNs (2 nmol/0.6 μl per side) into the bilateral rACC on S-CPA. Values are mean ± SEM time in the formalin-paired compartment on pre-conditioning and post-conditioning days (E), and CPA scores (F). *p
    Figure Legend Snippet: NMDA receptor NR2A and NR2B subunits fail to affect the induction of S-CPA A, Schematic of the protocol for behavioral testing. B,C, Histograms showing low-intensity (0.5 mA, 2 s) electric foot shock-induced CPA. Mean ± SEM time spent in the treatment-paired compartment on pre- and post-conditioning day (B). Mean ± SEM S-CPA scores, i.e. the time spent in the treatment-paired compartment on the pre-conditioning day minus that on the post-conditioning day (C). D, Schematic of the behavioral procedure used in subsequent studies. E,F, Histograms showing effects of microinjection of NR2B antagonist Ifenprodil (0.2 μg/μl, 0.6 μl per side), Ro 25-6981 (2 μg/μl, 0.6 μl per side), or NR2A AS ODNs (2 nmol/0.6 μl per side) into the bilateral rACC on S-CPA. Values are mean ± SEM time in the formalin-paired compartment on pre-conditioning and post-conditioning days (E), and CPA scores (F). *p

    Techniques Used:

    Related Articles

    Marker:

    Article Title: Epilepsy-associated GRIN2A mutations reduce NMDA receptor trafficking and agonist potency – molecular profiling and functional rescue
    Article Snippet: .. Proteins were then transferred onto Hybond-ECL nitrocellulose membrane (Amersham) which was cut on the 38 kDa marker and the top part ( > 38 kDa) incubated with anti-GluN2A antibody (M264, Sigma) at 1:1000 and the bottom part with anti-GAPDH antibody (AM4300, Ambion) 1:10,000. .. Primary antibodies were detected with horseradish peroxidase conjugated anti-rabbit (GluN2A) and anti-mouse (GAPDH) antibodies (Sigma) at 1:10,000 dilution, followed by signal development with SuperSignal West Dura reagent (ThermoFisher).

    Western Blot:

    Article Title: Rabphilin 3A retains NMDA receptors at synaptic sites through interaction with GluN2A/PSD-95 complex
    Article Snippet: .. Antibodies The following primary antibodies were used: rabbit anti-N-term GluN2A (480031, Invitrogen; dilution 1:100 ICC), rabbit anti-GluN2A (M264, Sigma-Aldrich; dilution 1:1,000 WB), mouse anti-N-term GluN2B (75–101, Neuromab; dilution 1:100 ICC, 1:1,000 WB), anti-PSD-95 (75–028; Neuromab; dilution 1:250 ICC, 1:1,000 WB) and anti-Pan-shank (75–089, Neuromab; dilution 1:200), rabbit anti-N-term GluA1 (MAB2263, Millipore; Billenca, MA; dilution 1:100 ICC), rabbit anti-Rph3A (NB100–92221, Novus biological; dilution 1:100 ICC), mouse anti-Rph3A (RM3701, ECM biosciences; dilution 1:100 ICC, 1:1,000 WB), rabbit anti-Rab8 (D22D8, Cell Signaling; dilution 1:1,000 WB), mouse anti-Meox2 (ab117551, Abcam; dilution 1:1,000 WB), mouse anti-Tubulin (T9026, Sigma-Aldrich; dilution 1:5,000 WB). .. The following secondary antibodies were used: goat anti-mouse-HRP (172–1,011) and goat anti-rabbit-HRP (172–1,019, Bio-Rad), goat anti-mouse-Alexa488 (A-11029), goat anti-mouse-Alexa555 (A-21422), goat anti-mouse-Alexa633 (A-21052), goat anti-rabbit-Alexa488 (A-11034) and goat anti-rabbit-Alexa555 (A-21429; Life Technologies).

    Article Title: High frequency stimulation induces sonic hedgehog release from hippocampal neurons
    Article Snippet: .. The anti-NR2A antibody (Cat. No. M264, 1:1000 dilution for western blots), anti-α-tubulin antibody (Cat. No. T6074, 1:3000 dilution for western blots) and anti-GAPDH antibody (Cat. No. G8795, 1:3000 dilution for western blots) were purchased from Sigma. .. The anti- axon-specific microtubule associated protein SMI312 antibody (Cat. No. SMI312R, 1: 200 dilution for immunostaining) was purchased from Covance, USA.

    Incubation:

    Article Title: Epilepsy-associated GRIN2A mutations reduce NMDA receptor trafficking and agonist potency – molecular profiling and functional rescue
    Article Snippet: .. Proteins were then transferred onto Hybond-ECL nitrocellulose membrane (Amersham) which was cut on the 38 kDa marker and the top part ( > 38 kDa) incubated with anti-GluN2A antibody (M264, Sigma) at 1:1000 and the bottom part with anti-GAPDH antibody (AM4300, Ambion) 1:10,000. .. Primary antibodies were detected with horseradish peroxidase conjugated anti-rabbit (GluN2A) and anti-mouse (GAPDH) antibodies (Sigma) at 1:10,000 dilution, followed by signal development with SuperSignal West Dura reagent (ThermoFisher).

    other:

    Article Title: Phosphoproteomic Alterations of Ionotropic Glutamate Receptors in the Hippocampus of the Ts65Dn Mouse Model of Down Syndrome
    Article Snippet: Antibodies Antibodies were obtained from commercial suppliers as it follows: anti-PSD95 (Neuromab, #P78352), anti-synaptophysin (Sigma-Aldrich, S5768), anti-β-actin (Sigma-Aldrich, A5316), anti-GluN2A (Sigma-Aldrich, M264), anti-GluN1 (Millipore, #AB9864R), anti-CaMKIIα (CST, #50049), anti-Fyn (CST, #4023), anti-pS890 GluN1 (CST, #3381), anti-pS1284 GluN2B (CST, #5355), anti-rabbit IgG-horseradish peroxidase (HRP; Dako, #P0448), and anti-mouse IgG-HRP (Dako, # P0447).

    Article Title: The STEP61 interactome reveals subunit-specific AMPA receptor binding and synaptic regulation
    Article Snippet: Mouse anti-STEP (cat. no. NB300-202; Novagen), rabbit anti-SynGAP (cat. no. 3200; Cell Signaling), rabbit anti–Kalirin-7 (laboratory made), mouse anti–NLGN-1 (cat. no. 129111; Synaptic Systems), rabbit anti-NBEA (cat. no. 194003; Synaptic Systems), rabbit anti-GluA1 (custom made), rabbit anti-GluA2/3 , mouse anti-GluA2 (clone L21/32; NeuroMab), rabbit anti-GluA2 (cat. 82103; Synaptic Systems), rabbit anti-GluA3 (cat. no. 3437; Cell Signaling), rabbit anti-GluN2A (cat. no. M264; Sigma), mouse anti-GluN2B (clone N59/36; NeuroMab), mouse anti-GluN1 , mouse anti–PSD-95 (clone K28/43; NeuroMab), mouse anti-SAP102 (clone N19/2; NeuroMab), rabbit anti-Fyn (cat. no. 4023; Cell Signaling), rabbit anti-Src (cat. no. 2110; Cell Signaling), mouse anti-GAPDH (cat. no. sc-365062; Santa Cruz), mouse anti-FLAG (cat. no. F1804; Sigma), rabbit anti-FLAG (cat. no. 7425; Sigma), rabbit anti-HA (cat. no. 3724; Cell Signaling), mouse anti-HA (cat. no. 2367; Cell Signaling), rabbit anti-myc (cat. no. 2278; Cell Signaling), rabbit anti-GST (cat. no. A190-122A; Bethyl Laboratories), mouse anti–β-actin (cat. no. G043; abm), and mouse anti-4G10 (cat. no. 05-321; Millipore).

    Immunocytochemistry:

    Article Title: Rabphilin 3A retains NMDA receptors at synaptic sites through interaction with GluN2A/PSD-95 complex
    Article Snippet: .. Antibodies The following primary antibodies were used: rabbit anti-N-term GluN2A (480031, Invitrogen; dilution 1:100 ICC), rabbit anti-GluN2A (M264, Sigma-Aldrich; dilution 1:1,000 WB), mouse anti-N-term GluN2B (75–101, Neuromab; dilution 1:100 ICC, 1:1,000 WB), anti-PSD-95 (75–028; Neuromab; dilution 1:250 ICC, 1:1,000 WB) and anti-Pan-shank (75–089, Neuromab; dilution 1:200), rabbit anti-N-term GluA1 (MAB2263, Millipore; Billenca, MA; dilution 1:100 ICC), rabbit anti-Rph3A (NB100–92221, Novus biological; dilution 1:100 ICC), mouse anti-Rph3A (RM3701, ECM biosciences; dilution 1:100 ICC, 1:1,000 WB), rabbit anti-Rab8 (D22D8, Cell Signaling; dilution 1:1,000 WB), mouse anti-Meox2 (ab117551, Abcam; dilution 1:1,000 WB), mouse anti-Tubulin (T9026, Sigma-Aldrich; dilution 1:5,000 WB). .. The following secondary antibodies were used: goat anti-mouse-HRP (172–1,011) and goat anti-rabbit-HRP (172–1,019, Bio-Rad), goat anti-mouse-Alexa488 (A-11029), goat anti-mouse-Alexa555 (A-21422), goat anti-mouse-Alexa633 (A-21052), goat anti-rabbit-Alexa488 (A-11034) and goat anti-rabbit-Alexa555 (A-21429; Life Technologies).

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 90
    Millipore rabbit monoclonal anti glun2a
    Decoration of <t>GluN1/GluN2A</t> NMDA receptors by Sig1Rs. A , The protein isolated from tsA 201 cells expressing Sig1R-V5, wild-type GluN1, and GluN2A-FLAG by anti-FLAG immunoaffinity chromatography was analyzed by immunoblotting using anti-V5 (left), anti-GluN1
    Rabbit Monoclonal Anti Glun2a, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit monoclonal anti glun2a/product/Millipore
    Average 90 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    rabbit monoclonal anti glun2a - by Bioz Stars, 2020-08
    90/100 stars
      Buy from Supplier

    91
    Millipore rabbit anti nr2a
    Expression of glutamate receptors. (A) Representative western-blot analysis of GluR1, <t>NR2A,</t> and NR2B subunits in the ACC homogenates. Levels of GluR1 (B) , NR2A (C), and NR2B (D) in saline control, vehicle-, T3-, and T4-treated HT mice. n = 5 in each group. * p
    Rabbit Anti Nr2a, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti nr2a/product/Millipore
    Average 91 stars, based on 14 article reviews
    Price from $9.99 to $1999.99
    rabbit anti nr2a - by Bioz Stars, 2020-08
    91/100 stars
      Buy from Supplier

    Image Search Results


    Decoration of GluN1/GluN2A NMDA receptors by Sig1Rs. A , The protein isolated from tsA 201 cells expressing Sig1R-V5, wild-type GluN1, and GluN2A-FLAG by anti-FLAG immunoaffinity chromatography was analyzed by immunoblotting using anti-V5 (left), anti-GluN1

    Journal: The Journal of Neuroscience

    Article Title: The ?-1 Receptor Interacts Directly with GluN1 But Not GluN2A in the GluN1/GluN2A NMDA Receptor

    doi: 10.1523/JNEUROSCI.3360-13.2013

    Figure Lengend Snippet: Decoration of GluN1/GluN2A NMDA receptors by Sig1Rs. A , The protein isolated from tsA 201 cells expressing Sig1R-V5, wild-type GluN1, and GluN2A-FLAG by anti-FLAG immunoaffinity chromatography was analyzed by immunoblotting using anti-V5 (left), anti-GluN1

    Article Snippet: The eluted samples were analyzed by SDS-PAGE, followed by immunoblotting using the following antibodies, as appropriate: mouse monoclonal anti-FLAG (clone M2; Sigma-Aldrich), mouse monoclonal anti-V5 (Invitrogen), mouse monoclonal anti-GluN1 (ab134308, raised against amino acids 42–361 of GluN1; Abcam), and rabbit monoclonal anti-GluN2A (clone A12W, raised against residues 1265–1464 of mouse GluN2A; Millipore).

    Techniques: Isolation, Expressing, Chromatography

    In situ proximity ligation assays for interactions of the Sig1R with either GluN1 or GluN2A. A , The tsA 201 cells were transfected individually with DNA encoding Sig1R-FLAG, GluN1-HA (ABD), GluN1-HA (post-TMD), or GluN2A-HA (ABD). Cells were permeabilized

    Journal: The Journal of Neuroscience

    Article Title: The ?-1 Receptor Interacts Directly with GluN1 But Not GluN2A in the GluN1/GluN2A NMDA Receptor

    doi: 10.1523/JNEUROSCI.3360-13.2013

    Figure Lengend Snippet: In situ proximity ligation assays for interactions of the Sig1R with either GluN1 or GluN2A. A , The tsA 201 cells were transfected individually with DNA encoding Sig1R-FLAG, GluN1-HA (ABD), GluN1-HA (post-TMD), or GluN2A-HA (ABD). Cells were permeabilized

    Article Snippet: The eluted samples were analyzed by SDS-PAGE, followed by immunoblotting using the following antibodies, as appropriate: mouse monoclonal anti-FLAG (clone M2; Sigma-Aldrich), mouse monoclonal anti-V5 (Invitrogen), mouse monoclonal anti-GluN1 (ab134308, raised against amino acids 42–361 of GluN1; Abcam), and rabbit monoclonal anti-GluN2A (clone A12W, raised against residues 1265–1464 of mouse GluN2A; Millipore).

    Techniques: In Situ, Ligation, Transfection

    Expression, isolation, and analysis of NMDA receptors. A–C , functional properties of wild-type and FLAG/His 8 -tagged GluN2A coexpressed with untagged GluN1 in Xenopus oocytes. A , currents activated by saturating concentrations of l -glutamate and glycine (100 μ m for both) in Xenopus oocytes expressing untagged GluN1 in combination with either wild-type (●; n = 9) or tagged (♦; n = 16) GluN2. The horizontal lines denote the means of the two distributions, which are not significantly ( NS ) different from each other ( p = 0.174, Student's t test). B , glycine concentration-response curves in the presence of 100 μ m glutamate. Curves were fitted with the Hill equation. EC 50 values were 1.7 ± 0.1 and 1.4 ± 0.1 μ m for wild-type and tagged GluN2A, respectively. Hill coefficients ( n H ) were 1.4 ± 0.1 and 1.4 ± 0.1 for wild-type and tagged GluN2A, respectively ( n = 3–4). C , glutamate concentration-response curves in the presence of 100 μ m glycine. EC 50 values were 3.9 ± 0.2 and 3.8 ± 0.2 μ m for wild-type and tagged GluN2A, respectively. n H values were 1.4 ± 0.1 and 1.6 ± 0.1 for wild-type and tagged GluN2A, respectively ( n = 3–4). D , samples of protein isolated from cotransfected HEK293T cells by affinity chromatography were analyzed by SDS-PAGE, followed by either silver staining ( left panel ) or immunoblotting using anti-GluN1 and anti-GluN2A antibodies ( right panel ). E , low-magnification AFM image of a sample of isolated NMDA receptors ( arrowheads ). Scale bar = 200 nm; shade-height scale = 0–5 nm. F , gallery of zoomed images of NMDA receptor particles that were decorated with an anti-GluN1 antibody ( arrowheads ). Scale bar = 20 nm; shade-height scale = 0–5 nm.

    Journal: The Journal of Biological Chemistry

    Article Title: Visualization of Structural Changes Accompanying Activation of N-Methyl-d-aspartate (NMDA) Receptors Using Fast-scan Atomic Force Microscopy Imaging *

    doi: 10.1074/jbc.M112.422311

    Figure Lengend Snippet: Expression, isolation, and analysis of NMDA receptors. A–C , functional properties of wild-type and FLAG/His 8 -tagged GluN2A coexpressed with untagged GluN1 in Xenopus oocytes. A , currents activated by saturating concentrations of l -glutamate and glycine (100 μ m for both) in Xenopus oocytes expressing untagged GluN1 in combination with either wild-type (●; n = 9) or tagged (♦; n = 16) GluN2. The horizontal lines denote the means of the two distributions, which are not significantly ( NS ) different from each other ( p = 0.174, Student's t test). B , glycine concentration-response curves in the presence of 100 μ m glutamate. Curves were fitted with the Hill equation. EC 50 values were 1.7 ± 0.1 and 1.4 ± 0.1 μ m for wild-type and tagged GluN2A, respectively. Hill coefficients ( n H ) were 1.4 ± 0.1 and 1.4 ± 0.1 for wild-type and tagged GluN2A, respectively ( n = 3–4). C , glutamate concentration-response curves in the presence of 100 μ m glycine. EC 50 values were 3.9 ± 0.2 and 3.8 ± 0.2 μ m for wild-type and tagged GluN2A, respectively. n H values were 1.4 ± 0.1 and 1.6 ± 0.1 for wild-type and tagged GluN2A, respectively ( n = 3–4). D , samples of protein isolated from cotransfected HEK293T cells by affinity chromatography were analyzed by SDS-PAGE, followed by either silver staining ( left panel ) or immunoblotting using anti-GluN1 and anti-GluN2A antibodies ( right panel ). E , low-magnification AFM image of a sample of isolated NMDA receptors ( arrowheads ). Scale bar = 200 nm; shade-height scale = 0–5 nm. F , gallery of zoomed images of NMDA receptor particles that were decorated with an anti-GluN1 antibody ( arrowheads ). Scale bar = 20 nm; shade-height scale = 0–5 nm.

    Article Snippet: Purified proteins were analyzed by SDS-PAGE and immunoblotting using mouse anti-GluN1 monoclonal antibody (clone 54.1, MAB363, Millipore) and rabbit anti-GluN2A monoclonal antibody (A12W, 04-901, Millipore).

    Techniques: Expressing, Isolation, Functional Assay, Concentration Assay, Affinity Chromatography, SDS Page, Silver Staining

    Expression of glutamate receptors. (A) Representative western-blot analysis of GluR1, NR2A, and NR2B subunits in the ACC homogenates. Levels of GluR1 (B) , NR2A (C), and NR2B (D) in saline control, vehicle-, T3-, and T4-treated HT mice. n = 5 in each group. * p

    Journal: Molecular Pain

    Article Title: Decreased pain threshold and enhanced synaptic transmission in the anterior cingulate cortex of experimental hypothyroidism mice

    doi: 10.1186/1744-8069-10-38

    Figure Lengend Snippet: Expression of glutamate receptors. (A) Representative western-blot analysis of GluR1, NR2A, and NR2B subunits in the ACC homogenates. Levels of GluR1 (B) , NR2A (C), and NR2B (D) in saline control, vehicle-, T3-, and T4-treated HT mice. n = 5 in each group. * p

    Article Snippet: Rabbit anti-NR2A and anti-NR2B were purchased from Millipore (Billerica, MA).

    Techniques: Expressing, Western Blot, Mouse Assay

    CD3ζ is required for the synaptic localization of GluN2A and the GluN2A–PSD95 interaction. A , Representative images of 15 DIV cultured cortical neurons from CD3ζ +/+ and CD3ζ −/− mice double labeled with GluN2A and Synapsin I. Scale bar, 10 μm. B , The number of GluN1, GluN2A, GluN2B, GluA1, and PSD95 clusters per length of dendrite was measured in 15 DIV CD3ζ +/+ and CD3ζ −/− cultured cortical neurons immunostained for each protein ( n = 3–4). C , Levels of synaptic GluN1, GluN2A, GluN2B, GluA1, and PSD95 clusters in 15 DIV CD3ζ +/+ and CD3ζ −/− cultured cortical neurons immunostained by double labeling of each postsynaptic protein with the presynaptic protein synapsin I. Data are mean ± SEM ( n = 3–4). *** p

    Journal: The Journal of Neuroscience

    Article Title: Impaired Spatial Memory in Mice Lacking CD3ζ Is Associated with Altered NMDA and AMPA Receptors Signaling Independent of T-Cell Deficiency

    doi: 10.1523/JNEUROSCI.3028-13.2013

    Figure Lengend Snippet: CD3ζ is required for the synaptic localization of GluN2A and the GluN2A–PSD95 interaction. A , Representative images of 15 DIV cultured cortical neurons from CD3ζ +/+ and CD3ζ −/− mice double labeled with GluN2A and Synapsin I. Scale bar, 10 μm. B , The number of GluN1, GluN2A, GluN2B, GluA1, and PSD95 clusters per length of dendrite was measured in 15 DIV CD3ζ +/+ and CD3ζ −/− cultured cortical neurons immunostained for each protein ( n = 3–4). C , Levels of synaptic GluN1, GluN2A, GluN2B, GluA1, and PSD95 clusters in 15 DIV CD3ζ +/+ and CD3ζ −/− cultured cortical neurons immunostained by double labeling of each postsynaptic protein with the presynaptic protein synapsin I. Data are mean ± SEM ( n = 3–4). *** p

    Article Snippet: The primary antibodies used in this study were as follows: mouse anti-EphA4 (1 μg/ml; Invitrogen), rabbit anti-CD3ζ (0.5 μg/ml; Spring Bioscience), mouse anti-PSD95 (1 μg/ml; Abcam), mouse anti-GluN1 (1 μg/ml; BD Pharmingen), rabbit anti-GluN2A (2 μg/ml; Millipore), mouse anti-GluN2B (2 μg/ml; NeuroMab), rabbit anti-GluA1 (5 μg/ml; Calbiochem), rabbit anti-Phospho831 GluA1 (Millipore), rabbit anti-Phospho286 CaMKII (1:200; Promega), mouse anti-CaMKIIα (0.5 μg/ml; Thermo Scientific), or mouse anti-GAPDH (0.2 μg/ml; Santa Cruz Biotechnologies).

    Techniques: Cell Culture, Mouse Assay, Labeling