rabbit anti nmdar 2b  (Alomone Labs)

Journal: Cell reports

Article Title: The exocyst subunit EXOC2 regulates the toxicity of expanded GGGGCC repeats in C9ORF72 -ALS/FTD

doi: 10.1016/j.celrep.2024.114375

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit anti-NMDAR 2B , Alomone Labs , Cat#AGC-003.

Techniques: Reverse Transcription, Enzyme-linked Immunosorbent Assay, Control, Recombinant, Plasmid Preparation, Software

Journal: Frontiers in Cellular Neuroscience

Article Title: Src Family Kinases Inhibition Ameliorates Hypoxic-Ischemic Brain Injury in Immature Rats

doi: 10.3389/fncel.2021.746130

Figure Lengend Snippet: Activated NR2B (Tyr1472) expression, the enhanced interaction of NR2B with Src, and changes in the levels of p-Src and p-NR2B after the administration of 4-amino-5-(4-chlorophenyl)-7-( t -butyl) pyrazolo [3,4- d ] pyrimidine (PP2) to the immature rat HI brain injury model. (A) Immunoblots showing the levels of multiple proteins. Primary antibodies are listed on the left. (B) Quantification of NR2B (Tyr1472) (p-NR2B) and NR2B levels at 2 h post injury (HI 2 h). N = 7 animals per group. (C) Coimmunoprecipitation was conducted using right hemisphere homogenates with the Src antibody or rabbit IgG as a control. Immune complexes were detected using immunoblotting (IB) with anti-Src and anti-NR2B antibodies. (D) Immunoblots showing the levels of multiple proteins. Primary antibodies are listed on the left. (E) Quantification of the levels of p-Src and Src at HI 2 h. N = 4–6 animals per group. (F) Quantification of the levels of p-NR2B and NR2B at HI 2 h. N = 4–6 animals per group. * p < 0.05 and ** p < 0.01.

Article Snippet: The blots were then blocked with 5% bovine serum albumin in TBS buffer for 1 h at room temperature and probed overnight at 4°C by incubation with the following primary antibodies: Src (1:1,000, CST, catalog number: #2123), phosphor-Src (Tyr 416, 1:1,000, CST, catalog number: #6943), NMDAR subunit NR2B (1:1,000, CST, catalog number: #14544), phospho-NR2B (Tyr 1472, 1:1,000, CST, catalog number: #4208), myelin basic protein (MBP) (1:1,000, Biolegend, SMI99), Iba-1 (1:1,000, Abcam, ab178847), glial fibrillary acidic protein (GFAP) (1:1,000, CST, 12389), β-actin (1:5,000, Absin, abs137975), and GAPDH (1:1,000, Absin, abs132004), followed by appropriate secondary HRP-conjugated antibodies (anti-rabbit, 1:5,000, Absin, abs20002; anti-mouse, 1:5,000, Absin, abs20001).

Techniques: Expressing, Western Blot, Control

Journal: The Journal of Pharmacology and Experimental Therapeutics

Article Title: Adenylyl Cyclase 1 Is Required for Ethanol-Induced Locomotor Sensitization and Associated Increases in NMDA Receptor Phosphorylation and Function in the Dorsal Medial Striatum

doi: 10.1124/jpet.117.242321

Figure Lengend Snippet: Repeated ethanol (EtOH) selectively enhanced GluN2B-NMDAR phosphorylation in the DMS in WT, but not AC1KO (KO) mice or mice pretreated with NB001 ( n = 3–5/group). Averaged data (left) and representative immunoblots compiled from groups of images from different parts of the same gel (right) are depicted for DMS (A), DLS (B), or NAc (C) lysates prepared following repeated saline (sal) or EtOH treatment and probed for total and pTyr-1472-GluN2B. (A) A significant increase in pGluN2B levels in the DMS of WT mice was observed following repeated EtOH treatment relative to saline-treated controls, which was absent in AC1KO and NB001 pretreated mice. (B) Repeated ethanol treatment decreased levels of pGluN2B in the DLS equivalently across WT, AC1KO, and NB001-treated mice. (C) No changes in pGluN2B levels in the NAc were observed in response to ethanol or absence of AC1 signaling. Total GluN2B levels were unaffected by AC1 status or ethanol treatment in all three regions. Significance determined by two-way ANOVA and Sidak’s post hoc test, * P < 0.05, compared with saline-treated controls; # P < 0.05, compared with WT EtOH.

Article Snippet: Equal amounts of protein (10 μ g) were separated using SDS-PAGE (4%–12% Bis-Tris NuPAGE gel [Invitrogen, Carlsbad, CA]), transferred to a nitrocellulose membrane, and probed with primary antibodies against pTyr-1472-GluN2B-NMDAR (1:2000, 4208; Cell Signaling Technology, Danvers, MA), GluN2B-NMDAR (1:2000, 4212; Cell Signaling Technology), and actin (1:5000, A5060; Sigma-Aldrich).

Techniques: Phospho-proteomics, Western Blot, Saline

Journal: The Journal of Pharmacology and Experimental Therapeutics

Article Title: Adenylyl Cyclase 1 Is Required for Ethanol-Induced Locomotor Sensitization and Associated Increases in NMDA Receptor Phosphorylation and Function in the Dorsal Medial Striatum

doi: 10.1124/jpet.117.242321

Figure Lengend Snippet: Ethanol (EtOH) upregulation of NMDAR-mediated transmission is absent in AC1KO (KO) mice. (A) Representative traces of NMDAR-mediated eEPSCs and (B) average NMDAR-mediated eEPSC amplitude (± S.E.M.) in WT and AC1KO mice repeatedly treated with saline (sal) or EtOH. EtOH produced an increase in NMDAR-mediated eEPSC amplitude only in WT mice (two-way ANOVA and Sidak’s post hoc test, ** P < 0.01). (C) Average NMDAR-mediated eEPSC amplitude before (BL) and after (+Ro) bath application of the GluN2B selective antagonist Ro 25-6981 (0.5 µ M) in WT mice. EtOH-induced upregulation of NMDAR-mediated transmission involved an increased contribution of GluN2B-containing NMDARs (two-way RM ANOVA and Sidak’s post hoc test, * P < 0.05). (D) Average central location of medium spiny neuron recordings was 1.10 mm from bregma (± 0.5 mm).

Article Snippet: Equal amounts of protein (10 μ g) were separated using SDS-PAGE (4%–12% Bis-Tris NuPAGE gel [Invitrogen, Carlsbad, CA]), transferred to a nitrocellulose membrane, and probed with primary antibodies against pTyr-1472-GluN2B-NMDAR (1:2000, 4208; Cell Signaling Technology, Danvers, MA), GluN2B-NMDAR (1:2000, 4212; Cell Signaling Technology), and actin (1:5000, A5060; Sigma-Aldrich).

Techniques: Transmission Assay, Saline, Produced