rabbit polyclonal anti nr2a  (Cell Signaling Technology Inc)

Journal: Frontiers in Cellular Neuroscience

Article Title: Rescue of sharp wave-ripples and prevention of network hyperexcitability in the ventral but not the dorsal hippocampus of a rat model of fragile X syndrome

doi: 10.3389/fncel.2023.1296235

Figure Lengend Snippet: Western blotting analysis of the three NMDAR subunits, NR1, NR2A, and NR2B in WT and KO dorsal (A) and ventral hippocampus (B) . The results of the statistical analysis (independent t -test) showed similar expressions between the WT and KO dorsal hippocampus (NR1: t 14 = 0.107, p = 0.813; NR2A: t 14 = –0.24, p = 0.369; NR2B: t 14 = 1.128, p = 0.842; n = 8 rats in either genotype), and ventral hippocampus (NR1: t 14 = –0.241, p = 0.813; NR2A: t 14 = 0.803, p = 0.435; NR2B: t 14 = 0.203, p = 0.842; n = 8 rats in either genotype). Also, the NR2A/NR2B ratio is similar in WT and KO dorsal ( t 10.15 = –1.38, p = 0.199) and ventral hippocampus ( t 9.72 = 0.383, p = 0.71). The expression of NR2A subunit and the NR2A/NR2B ratio is higher in the dorsal compared with the ventral hippocampus in both WT (NR2A: t 14 = 4.27, p < 0.001, and NR2A/NR2B ratio: t 14 = 3.08, p = 0.008) and KO rats (NR2A: t 14 = 4.017, p = 0.001, and NR2A/NR2B ratio: t 14 = 3.01, p = 0.009). In contrast, NR1 and NR2B are similarly expressed in WT and KO dorsal (NR1: t 14 = 1.308, p = 0.212, and NR2B: t 14 = 1.09, p = 0.294) and ventral hippocampus (NR1: t 14 = 1.039, p = 0.316, and NR2B: t 14 = 0.187, p = 0.854). (C) Images of individual western blot samples with detected bands of the NMDA receptor protein subunits, and the corresponding loading marker band of beta actin. “ns” denotes not significant difference.

Article Snippet: Membranes were next incubated overnight at 4°C with the following primary antibodies diluted in 3% PBST: rabbit anti-α1 GABA A R polyclonal antibody (1:2500 #06-868, Millipore Sigma), rabbit anti-NR1 monoclonal antibody (1:1000 #D65B7, Cell Signaling), rabbit anti-NR2A polyclonal antibody (1:1000 #4205, Cell Signaling), rabbit anti-NR2B monoclonal antibody (1:1000 #B8E10, Cell Signaling) and rabbit anti-β-actin polyclonal antibody (1:15000 #E-AB-20058, Elabscience).

Techniques: Western Blot, Expressing, Marker

Journal: BMC Neuroscience

Article Title: Expression of AMPA and NMDA receptor subunits in the cervical spinal cord of wobbler mice

doi: 10.1186/1471-2202-7-71

Figure Lengend Snippet: ionotropic glutamate receptors in the lamina IX in the cervical spinal spinal cord of four-week-old wobbler mice . UPPER PANELS. Representative photomicrographs showing the patterns of GFAP immunostaining in the cervical spinal cord region of 12-week-old mice (A) and age-matched symptomatic wobbler mice (B). CENTRAL PANELS (C-V). Triple staining experiments using Nissl (C, G, K, O, S, purple colour) GFAP (D, H, L, P, T, red colour) and GluR 1–4 or NR2A (E, I, M, Q, U, green colour) in the anterior horn of four-week-old wobbler mice. The merge between the three different colours is shown in panels F, J, N, R, V. LOWER PANELS (W-Z). Triple staining showing Nissl (W), GFAP (X), GluR4 (Y) and the merge between the three different colurs (Z), in the white matter in the anterior region of the cervical spinal cord, four-week-old wobbler mouse. Scale bar, A-B, 100 μm. C-V, 30 μm. W-Z 40 μm.

Article Snippet: For the NMDA receptor we used an anti-mouse monoclonal NR1 antibody (Catatolg No 32-0500, Zymed, Invitrogen, Carlsbad, CA, US; 1:100) and an anti-rabbit polyclonal NR2A antibody (Molecular Probes, Invitrogen, Carlsbad, CA, US; 1:100).

Techniques: Immunostaining, Staining

Journal: BMC Neuroscience

Article Title: Expression of AMPA and NMDA receptor subunits in the cervical spinal cord of wobbler mice

doi: 10.1186/1471-2202-7-71

Figure Lengend Snippet: NMDA receptor subunits in spinal cord of wobbler mice . Representative photomicrographs showing NR1 (A,C) and NR2A (B,D) immunostaining in the cervical spinal cord of four-week-old wobbler mice (B). Scale bar, A, B 100 μm. C, D 20 μm.

Article Snippet: For the NMDA receptor we used an anti-mouse monoclonal NR1 antibody (Catatolg No 32-0500, Zymed, Invitrogen, Carlsbad, CA, US; 1:100) and an anti-rabbit polyclonal NR2A antibody (Molecular Probes, Invitrogen, Carlsbad, CA, US; 1:100).

Techniques: Immunostaining

Journal: BMC Neuroscience

Article Title: Expression of AMPA and NMDA receptor subunits in the cervical spinal cord of wobbler mice

doi: 10.1186/1471-2202-7-71

Figure Lengend Snippet: western blotting analysis and quantification of the percentage of AMPA and NMDA receptor subunits expression in spinal cord of wobbler mice . Representative immunoblot from cervical spinal cord samples from 12-week-old healthy mice and age-matched wobbler mice. Left column: whole spinal cord homogenates. Right column: spinal cord TIF. Quantification of the mean values of immunodensity for CaMK, AMPA and NMDA receptor subunits, obtained from the cervical spinal wobbler mice in whole spinal cord homogenates (Homo) and in TIF. Data are representative of two independent experiments on two different homogenate preparations of four pooled animals for each group and replicated two times in each homogenate preparation. Values reported represent the percentage of the mean values of immunodensity obtained in 12-week-old wobbler mice compared to the levels measured in healthy littermates and normalized to 100. Both preparations samples were analyzed by Western blot analysis with CaMKII, GluR1, GluR2, GluR3, GluR4, NR1, and NR2A antibodies. The same amount of protein was loaded per lane.

Article Snippet: For the NMDA receptor we used an anti-mouse monoclonal NR1 antibody (Catatolg No 32-0500, Zymed, Invitrogen, Carlsbad, CA, US; 1:100) and an anti-rabbit polyclonal NR2A antibody (Molecular Probes, Invitrogen, Carlsbad, CA, US; 1:100).

Techniques: Western Blot, Expressing

Journal: PLoS ONE

Article Title: Brimonidine Blocks Glutamate Excitotoxicity-Induced Oxidative Stress and Preserves Mitochondrial Transcription Factor A in Ischemic Retinal Injury

doi: 10.1371/journal.pone.0047098

Figure Lengend Snippet: BMD or vehicle were injected once by IP at 24 hours before and at the time of initial IOP elevation for 24 hours after ischemia. (A) Vehicle-treated ischemic retina significantly increased GFAP protein expression compared with control retina. In contrast, BMD significantly decreased GFAP protein expression in ischemic retina. (B–E) GFAP immunohistochemisty. When the primary antibody for GFAP was omitted, there was no labeling of the secondary antibody (B). In comparison with control retina (C), vehicle-treated ischemic retina showed activation of müller cells (arrowheads) and astrocytes (D). In contrast, BMD significantly decreased activation of müller cells and astrocytes (E). ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer. Scale = 20 µm. (F) Vehicle-treated ischemic retina significantly increased NR1, NR2A and SOD2 protein expression compared with control retina. In contrast, BMD significantly decreased NR1, NR2A and SOD2 protein expression in ischemic retina. Values are mean ± SD (n = 4 retinas/group). *Significant at p<0.05 compared with non-ischemic contralateral control retina or vehicle-treated ischemic retina.

Article Snippet: The membrane was blocked with 5% nonfat dry milk and 0.1% Tween-20 in PBS, incubated with monoclonal mouse anti-GFAP (1∶3000; Sigma), goat polyclonal anti-Tfam antibody (1∶1000; Santa Cruz Biotechnology), mouse monoclonal anti-total OXPHOS complex antibody (containing a mixture of antibodies to COXI-IV and ATP synthase, 1∶3000; Invitrogen), rabbit polyclonal anti-Bax antibody (1∶500; Santa Cruz Biotechnology), rabbit polyclonal anti-Bcl-xL antibody (1∶1000; Cell Signaling, Danvers, MA), mouse monoclonal anti-phosphorylated Bad (pBad, 1∶2000; Cell Signaling), mouse monoclonal anti-NR1 antibody (1∶1000; BD Pharmingen, San Diego, CA), rabbit polyclonal anti-NR2A antibody (1∶500; Millipore), rabbit polyclonal anti-voltage-dependent anion channel (VDAC) antibody (Porin, 1∶1000; Calbiochem, La Jolla, CA) and mouse monoclonal anti-actin antibody (1∶5000, Millipore, Billerics, MA).

Techniques: Injection, Expressing, Labeling, Activation Assay