A-6473

antibody from Invitrogen Antibodies

Targeting: GRIN2A GluN2A, NMDAR2A

Provider product page for A-6473

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [19]
Comments [0]
Validations
Immunohistochemistry [2]
Other assay [14]

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Product number: A-6473 - Provider product page
Provider: Invitrogen Antibodies
Product name: NMDAR2A Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Other
Description: This antibody recognizes the 180 kDa 2A subunit from rat, mouse and human, and does not cross-react with the 2B or 2C subunits.

GRIN2A protein structure - A-6473 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 3 Expression of NMDAR subunits NR2A and NR2B, and MAGUKs PSD95 and SAP102, at different stages of development. A , Total protein extracts were obtained from hippocampal neurons at 2, 5, 7, 12, 15, 18, and 20 DIV and immunoblotted with antibodies against NR2B, NR2A, PSD95, SAP102, or N-cadherin as loading control. B , Results of qRT-PCR run with primers designed to determine mRNA levels for NR2B, NR2A, PSD95, or SAP102 at the same developmental stages; results were normalized against levels of GAPDH mRNA. Note that NR2B expression decreases with development, while NR2A and PSD95 increases. Expression of SAP102 remains relatively stable at these developmental stages. C , Immunodetection of endogenous NR2B, NR2A, PSD95, or SAP102 (green) and co-localization with presynaptic marker Bassoon or Piccolo (red) at 5 and 20 DIV. Insets show magnified views of boxed areas showing examples of synaptic clusters located in close apposition to presynaptic clusters.

Supportive validation

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Experimental details: Figure 6 Interaction of PSD95 with NMDAR subunits and PMCA isoforms. Brain lysates were incubated with anti-PSD95 antibodies conjugated to sepharose beads. Immunocomplexes were recovered and resolved by SDS-PAGE and the membranes were next probed with anti-NMDAR1 (A) , anti-NMDAR2A (B) , anti-NMDAR2B (C) , 5F10 (D) , anti-PMCA2 (E) or anti-PMCA4 (F) antibodies. Negative controls included sepharose-linked secondary antibodies (Control Ab) or sepharose beads only. PSD95 was also immunoprobed in beads-unbound fraction (unbound). Representative blots are shown. The bands intensity were densitometically quantified and the results were expressed as AU defined as the optical density per mg of protein (OD/mg protein). * P < 0.05 ketamine treated vs. saline, n = 5. CX, cortex; CB, cerebellum; H, hippocampus; ST, striatum.

Supportive validation

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Experimental details: Figure 5 Western blot analysis of PSD95 and N-methyl-D-aspartic acid receptor (NMDAR) subunit protein level in synaptosomal membranes. The protein level of PSD95, NMDAR1, NMDAR2A and NMDAR2B was determined by immunoblotting (A) and the bands intensity corresponding to the target protein was densitometrically analyzed (B) . The results are expressed as AU obtained following normalization to endogenous Na + /K + -ATPase level. Representative blots are shown. * P < 0.05 ketamine treated vs. saline, n = 5. CX, cortex; CB, cerebellum; H, hippocampus; ST, striatum.

Supportive validation

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Experimental details: Figure 4 Neto1 Binds to NMDA Receptors Independently of the C-Terminal PDZ Ligand (A, B) Immunoblots of immunoprecipitates from transfected HEK293 cell lysates. The transfected cDNAs are shown above each lane. CSF-1 EC-eGFP encodes the extracellular domain of CSF-1 fused to eGFP. Nrpn1 CUB12-eGFP encodes the two CUB domains from neuropilin-1 fused to eGFP. Blot, antibody used for immunoblot analysis; IP, antibody used for immunoprecipitation.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 6 Newly synthesized mEos3.2-KIF17 moves along the dendrites and colocalizes with NR2B. ( A to D ) Time-lapse images of newly synthesized mEos3.2-KIF17(+) (A), mEos3.2-KIF17(DeltaM) (B), and mEos3.2-KIF17(DeltaT) (C) along dendrites and their kymographs (D). Hippocampal neurons (DIV 17 to 19) transfected with mEos3.2-KIF17(+)3'UTR, mEos3.2-KIF17(DeltaM)3'UTR, and mEos3.2-KIF17(DeltaT)3'UTR were stimulated with NMDA (20 muM) after photoconversion of mEos3.2. Yellow arrowheads in (A) to (C), newly synthesized mEos3.2 fused to each construct of KIF17. Black arrowheads in (D), the time and location of KIF17 3'UTR-mediated local synthesis. Red lines in (A) to (C) trace the morphology of the dendrites. Scale bars, 5 mum. See also fig. S8. ( E and F ) Quantifications as average time when mEos3.2 fused to each construct of KIF17 was synthesized from NMDA bath application (E) and velocities for movements of mEos3.2 fused to each construct of KIF17 (F). P >= 0.05 (E), P < 0.001 (F), one-way ANOVA; *** P < 0.001, Bonferroni's post hoc comparison (F). n = 7 moving puncta from six culture neurons. ( G to I ) Representative images of dendrites expressing mEos3.2-KIF17(+) or mEos3.2-KIF17(DeltaT), which were immunostained with anti-NR2B or anti-NR2A antibodies (G), and comparisons of the colocalization of the anti-NR2B-positive puncta (H) and that of mEos3.2-KIF17(+) (I) in dendrites. Scale bar, 5 mum. *** P < 0.001, two-tailed t test. n = 7 dendrites from seven culture neurons.

Supportive validation

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Experimental details: Figure 4 Effect of repeated mephedrone or amphetamine administration during adolescence on MMP-9 ( A ) and NMDA subunits (GluN1, GluN2A, GluN2B) and PSD-95 ( B ) expression in the prefrontal cortex (PC) and hippocampus (HC) of rats (PND 87) that underwent the Barnes maze task. Representative blots for significant changes are presented, as well as full membranes are presented in the supplementary material (Figure S1) . Data are expressed as mean +- SEM. n = 8 rats/group. * p < 0.05 vs vehicle (0.9% NaCl). V--vehicle, M--mephedrone, A--amphetamine.

Supportive validation

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Experimental details: Figure 3 Changes in the expression of NMDA receptor subunits (GluN1, GluN2A and GluN2B) in the nucleus accumbens in rats following cocaine self-administration. All data are expressed as % of control (mean +- SEM). N = 6 rats/group. * p < 0.05 vs. yoked saline. ( a ), GluN1 protein levels [% of control]; ( b ), GluN2A protein levels [% of control]. ; ( c ), GluN2B protein levels [% of control].

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 3 Changes in the expression of GluN2A subunit in the PSD fraction of PLC and dHIP in rats following cocaine abstinence with extinction training a . Full membranes for GluN2A subunit are presented b . PLC-prelimbic cortex, dHIP-dorsal hippocampus, PSD-postsynaptic density, YS-yoked saline, YC-yoked cocaine, SA-cocaine self-administration. All data are expressed as mean +- SEM. N = 8 rats/group. * p < 0.05 vs. yoked saline

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 Western blot analysis of NMDAR subunit expression in CA1 region of hippocampus. The blots illustrate expression of ( A ) GluN2B, ( B ) GluN2A, and PSD95. ( C ) The bars represent the mean (+- SEM) ratio of expression for vehicle ( n = 4) and LPS treated ( n = 4) animals.