rabbit polyclonal anti nr2a  (Cell Signaling Technology Inc)

Journal: Frontiers in Endocrinology

Article Title: Dorzagliatin shows potential in preventing cognitive impairment in diabetes: evidence from Mendelian randomization analysis and animal study

doi: 10.3389/fendo.2025.1755359

Figure Lengend Snippet: Dorzagliatin prevented diabetes-induced downregulation of synaptic proteins in Goto Kakizaki rats. (A) Western blot analysis of selected glutamate receptors and postsynaptic density protein 95 (PSD-95) in hippocampus of each group. GAPDH was used as an internal control. (B–D) Statistics of GluN1 (B) , GluN2A (C) and PSD-95 (D) protein levels in the hippocampus of T2D (Goto Kakizaki-vehicle) and control (Wistar-vehicle). (E) Western blot analysis of selected glutamate receptors and postsynaptic density protein 95 (PSD-95) in the hippocampus of Goto Kakizaki-vehicle and Goto Kakizaki-dorzagliatin rats. (F–H) Statistics of GluN1 (F) , GluN2A (G) and PSD-95 (H) protein levels in the hippocampus of Goto Kakizaki-vehicle group and Goto Kakizaki-dorzagliatin group. Data are expressed as mean ± SEM. Student’s t test, two tailed. *P < 0.05, **P < 0.01. (n=4 per group). hippo, hippocampus; dorza, dorzagliatin.

Article Snippet: The membranes were blocked with 1×TBST and 5% BSA (tank blotting) or 5% skim milk (Semi-dry blotting) for 1 hour at room temperature and then incubated overnight at 4°C with following primary antibodies, respectively: mouse monoclonal anti-GluN1 antibody (Millipore, Cat. No: 05-432); rabbit polyclonal anti-GluN2A antibody (NOVUS, Cat. No: NB300-105); rabbit monoclonal anti-GLUT1 antibody (Abcam, Cat. No: ab115730); rabbit polyclonal anti-GLUT3 antibody (Bioss, Cat. No: bs-1207R); rabbit polyclonal anti-IR antibody (Abcam, Cat. No: ab137747); rabbit monoclonal anti-PSD95 antibody (Abcam, Cat. No: ab238135).

Techniques: Western Blot, Control, Two Tailed Test

Journal: iScience

Article Title: Pyramidal and parvalbumin neurons modulate the process of electroacupuncture stimulation for stroke rehabilitation

doi: 10.1016/j.isci.2024.109695

Figure Lengend Snippet: The alteration of neuronal activity during EA-mediated effect might be associated with the function of NMDARs (A) Representative raw glutamate signal traces (ΔF/F). Scale bar, ΔF/F: 1%, time: 2 s. (B) The glutamate level does not change significantly before and after EA in stroke-surgery mice following grip stimulus ( n = 8 mice per group). (C and D) AMPAR/NMDAR ratio was decreased at stroke 1D but not in the stroke 7D, and it does not change after EA stimulation ( n = 9 cells from 3 Sham+shamEA/Sham+EA mice, n = 10 cells from 3 Stroke+shamEA/Stroke+EA mice). (E and F) The amplitude of mEPSCs was increased at stroke 1D but not in the stroke 7D, and it does not happen after EA stimulation ( n = 12 cells from 3 Sham+shamEA/Sham+EA mice, n = 11 cells from 3 Stroke+shamEA/Stroke+EA mice). (G and H) Representative western blot and NR2B in the contralesional M1FL increased after stroke 1D, while acute EA could restore its expression level ( n = 6 mice per group). (I and J) Representative western blot and NR2A in the contralesional M1FL decreased after stroke 7D, and chronic EA could restore its expression level ( n = 6 mice per group). (C to J) One-way analysis of variance (ANOVA) with Tukey post hoc test. (B) Two-way analysis of variance (ANOVA ) with Tukey post hoc test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Recordings were performed on mice during the 9-11th week after birth. All data are represented by mean±SEM .

Article Snippet: Rabbit polyclonal Antibody to GluN2A , Invitrogen , Cat# A-6473; RRID: AB_1501807.

Techniques: Activity Assay, Western Blot, Expressing

Journal: iScience

Article Title: Pyramidal and parvalbumin neurons modulate the process of electroacupuncture stimulation for stroke rehabilitation

doi: 10.1016/j.isci.2024.109695

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal Antibody to GluN2A , Invitrogen , Cat# A-6473; RRID: AB_1501807.

Techniques: Virus, Recombinant, Blocking Assay, Software

Journal: Cell reports

Article Title: TRPM2 enhances ischemic excitotoxicity by associating with PKCγ

doi: 10.1016/j.celrep.2024.113722

Figure Lengend Snippet: (A) Graphic illustration showing how TAT-M2PBM works. The PKCγ binding motif of TRPM2 (M2PBM) in conjugation with the cell-penetrating peptide TAT was synthesized. TAT-M2PBM binds to the TRPM2 binding site for PKCγ, thus achieving the competitive inhibition on the binding of TRPM2 to PKCγ. (B) Co-immunoprecipitation of PKCγ by TRPM2 in HEK293T cells co-expressed with PKCγ and TRPM2 treated with TAT-SC (scramble) or TAT-M2PBM at 1 μM for 2 h. (C and D) Whole-cell current recording of TRPM2 in HEK293T cells transfected with PKCγ and TRPM2 treated with TAT-SC (green) or TAT-M2PBM (red). (C) Representative traces. PMA (10 μM) was used to induce TRPM2 activation, NMDG to test seal tightness, and ACA to block TRPM2 current. (D) Quantification of current amplitude (n = 9, 9). (E and F) CKAR real-time imaging in HEK293T cells transfected with PKCγ and TRPM2 treated with TAT-SC (green) or TAT-M2PBM (red). (E) Averaged representative traces from 5 randomly chosen cells. (F) Quantification of FRET changes (n = 10–20). (G and H) Whole-cell current recording of NMDARs in HEK293T cells transfected with NMDARs/PKCγ and TRPM2 treated with TAT-SC (green) or TAT-M2PBM (red). (G) Representative traces. NMDA (100 μM) was used to induce NMDAR activation. (H) Quantification of current amplitude (n = 11, 10, 11). (I and J) Whole-cell current recording of NMDARs in neurons isolated from the WT and M2KO mice treated with TAT-SC or TAT-M2PBM. (I) Representative traces. (J) Quantification of current amplitude (n = 15, 11, 9, 6). (K and L) Surface expression of GluN2a and GluN2b in HEK293T cells transfected with NMDAR/PKCγ and TRPM2 treated with TAT-SC or TAT-M2PBM. (K) Representative western blot (WB) bands. (L) Quantification of relative expression normalized to pan-cadherin (n = 6/group). (M–O) Whole-cell current recording of NMDARs in neurons isolated from the WT and M2KO mice treated with TAT-SC or TAT-M2PBM. (M) Representative traces. PMA at 10 μM was used to enhance NMDAR’s activity for 60 s. (N) Quantification of current amplitude before and after PMA perfusion. (O) Quantification of current increases after PMA perfusion (n = 6–10/group). ns, no statistical significance, *p < 0.05, **p < 0.01, ***p < 0.001; unpaired t test; mean ± SEM.

Article Snippet: Rabbit polyclonal antibodies to TRPM2 (Novus, NB110–81601, 1:50 in protein extraction for IP); Rabbit polyclonal antibodies to GluN2A (Cell Signaling Technology, 4205S, 1:1000 in 5% BSA for WB).

Techniques: Binding Assay, Conjugation Assay, Synthesized, Inhibition, Immunoprecipitation, Transfection, Activation Assay, Blocking Assay, Imaging, Isolation, Expressing, Western Blot, Activity Assay

Journal: Cell reports

Article Title: TRPM2 enhances ischemic excitotoxicity by associating with PKCγ

doi: 10.1016/j.celrep.2024.113722

Figure Lengend Snippet: (A) Co-immunoprecipitation of PKCγ, GluN2a, and GluN2b by TRPM2 in the brain from WT mice 2, 12, and 24 h after injection with TAT-SC (scramble) or TAT-M2PBM at 100 nmol/kg. (B) Injection strategy for evaluating short-term protective effects. (C and D) Brain injury in WT and Trpm2 deletion (M2KO) mice injected with TAT-SC or TAT-M2PBM (100 nmol/kg) 24 h after MCAO (n = 8, 7, 5, 5). (C) Triphenyl tetrazolium chloride (TTC) staining showing infract area (white). (D) Quantification of brain infarction and neurological deficit score. (E and F) Brain injury in WT mice injected with TAT-SC, TAT-EE 3 , or TAT-M2PBM 24 h after MCAO (n = 7, 9, 9). (E) TTC staining showing infract area (white). (F) Quantification of brain infarction and neurological deficit score. (G) Injection strategy for evaluating short-term protective effects. (H–K) Brain injury in WT and M2KO mice injected with TAT-SC or TAT-M2PBM 7 days after MCAO (n = 8, 9). (H) TTC staining showing infract area (white). (I and J) Quantification of brain infarction and neurological deficit score. (K) Quantification of rotarod test. ns, no statistical significance, *p < 0.05, **p < 0.01, ***p < 0.001; unpaired t test; mean ± SEM; scale bar: 5 mm.

Article Snippet: Rabbit polyclonal antibodies to TRPM2 (Novus, NB110–81601, 1:50 in protein extraction for IP); Rabbit polyclonal antibodies to GluN2A (Cell Signaling Technology, 4205S, 1:1000 in 5% BSA for WB).

Techniques: Immunoprecipitation, Injection, Staining

Journal: Cell reports

Article Title: TRPM2 enhances ischemic excitotoxicity by associating with PKCγ

doi: 10.1016/j.celrep.2024.113722

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal antibodies to TRPM2 (Novus, NB110–81601, 1:50 in protein extraction for IP); Rabbit polyclonal antibodies to GluN2A (Cell Signaling Technology, 4205S, 1:1000 in 5% BSA for WB).

Techniques: Virus, Recombinant, Sequencing, Bicinchoninic Acid Protein Assay, Isolation, Membrane, Extraction, Protein Extraction, Knock-Out, Mutagenesis, Subcloning, Plasmid Preparation, Software