mouse anti-nr2a  (Santa Cruz Biotechnology)

Journal: bioRxiv

Article Title: Identifying the function of the NMDA NR1 C2 subunit through its interaction with Magi-2 during inflammatory pain

doi: 10.1101/2024.01.30.578033

Figure Lengend Snippet: A ) Western analysis of NR1, NR2A, and NR2B NMDA receptor levels from spinal cord dorsal horn lysates of WT, heterozygous and homozygous ΔMagi-2 mice with actin as a loading control. B ) Densitometric analysis of 2A. Relative protein levels of each NMDA receptor subunit were compared using one-way ANOVA with Tukey post hoc analysis ( NR1 : WT vs. Het p=0.0249*; WT vs ΔMagi-2 p = 0.0004***) C) Immunoblot of NR1 following Magi-2 co-immunoprecipitation from dorsal horn lysates. D & F) Representative NMDA-evoked current traces recorded from lamina 1 neurons of WT, heterozygous, and ΔMagi-2 mice. Neurons were held at D) +60mV and F ) +40mV and then 50uM NMDA was bath applied for 60 seconds followed by a washout for 3 minutes. E & G) Current densities (peak NMDA-evoked current amplitude divided by cell capacitance compared by One-way ANOVA with Tukey post-hoc test. E ) +60mV: WT vs ΔMagi-2 p= 0.0392* G ) +40mV: WT vs. ΔMagi-2 p=0.0436*: one-way ANOVA with Tukey post hoc test.

Article Snippet: The membranes were then blocked in 5% milk in Tris Buffered Saline with 1% Tween (TBS-T) overnight at 4° C before being probed overnight at 4° C with Rabbit anti Magi-2 (1:1000, Sigma M4221); Mouse anti-NR1 (1:1000, Abcam ab134308); Mouse anti-NR2A (1:1000, Abcam ab124913); Rabbit anti-NR2B (1:1000, Abcam ab254356); Rabbit anti-NR1-C2 (1:1000, PhosphoSolutions 1506-C2); Rabbit anti-NR1-C2’ (1:1000, PhosphoSolutions 1507-C2’); or Rabbit anti-actin (1:5000, Sigma a2066).

Techniques: Western Blot, Immunoprecipitation

Journal: PLoS ONE

Article Title: Identification of CHIP as a Novel Causative Gene for Autosomal Recessive Cerebellar Ataxia

doi: 10.1371/journal.pone.0081884

Figure Lengend Snippet: ( A ) The expression of CHIP in the cerebellum (Cb), hippocampus (Hip), cerebral cortex (Ctx), pons (PN) and medulla oblongata (MO) of mouse brain as analyzed with immunohistochemistry. ( B ) Co-localization of CHIP (red) and calcium-binding protein calbindin D-28K (green) in Purkinje cells. ( C ) Co-localization of CHIP (red) and NR2A (green) in Cb, PN and MO. ( D ) Coexpression of flag-tagged WT, but not ARCA-associated CHIP mutants (CHIP N130I , CHIP W147C , CHIP L165F , CHIP Y207X , CHIP S236T ), with HA-Fbx2 promoted the degradation of NR2A. Expression vectors for CHIP, Fbx2 and NR2A were transfected into Human Embryonic Kidney 293 cells. At 36 h after transfection, cells were treated with cycloheximide (CHX, 100 μg/ml) and chased for different time periods. NR2A was detected with western blot using the Myc antibody. Quantitative analysis was performed using NIH ImageJ analysis software. Values represent the mean ± S.D. of three independent experiments.

Article Snippet: For the double immunofluorescence staining, the brain sections were treated with the rabbit anti-CHIP antibody and a mouse anti-NR2A monoclonal antibody (Millipore, USA).

Techniques: Expressing, Immunohistochemistry, Binding Assay, Transfection, Western Blot, Software

Journal: iScience

Article Title: Prkn knockout mice show autistic-like behaviors and aberrant synapse formation

doi: 10.1016/j.isci.2022.104573

Figure Lengend Snippet:

Article Snippet: Mouse anti-NR2A (1:1000) , EMD Millipore , Cat. #07-632; RRID: AB_310837.

Techniques: Software

Journal: Radiation Oncology (London, England)

Article Title: Whole-brain irradiation differentially modifies neurotransmitters levels and receptors in the hypothalamus and the prefrontal cortex

doi: 10.1186/s13014-020-01716-y

Figure Lengend Snippet: Expression of GABAa ( a ) GABAb ( b ), and NR1 subunit of the NMDA receptor ( c ). The protein expression was analyzed in the rat hypothalamus by Western Blott 1 month after sham (white bars) or whole-brain irradiation with a single dose of 11 Gy (black bars). The expression of the NR2A subunit was below standardized detection (data not shown). Data are expressed as means ± SEM. * p < 0.05 compared with the sham group using the Mann–Whitney U test

Article Snippet: Next, membranes were incubated overnight at 4 °C with either anti-GABAb mouse monoclonal antibody (1:1000, SC-166408, Santa Cruz, USA), anti-GABAa mouse monoclonal antibody (1:1000, sc-376282, Santa Cruz, USA), anti-NR1 monoclonal mouse antibody (1:200, BML-SA493-0015, Enzo, USA) or anti-NR2A monoclonal mouse antibody (1:200, sc-31540, Santa Cruz, USA) diluted in TBST containing 5% nonfat dry milk.

Techniques: Expressing, Western Blot, Irradiation, MANN-WHITNEY

Journal: Radiation Oncology (London, England)

Article Title: Whole-brain irradiation differentially modifies neurotransmitters levels and receptors in the hypothalamus and the prefrontal cortex

doi: 10.1186/s13014-020-01716-y

Figure Lengend Snippet: Expression of GABAa ( a ), GABAb ( b ), NR1 ( c ), and NR2A subunit of the NMDA receptor ( d ). The protein expression was analyzed in the rat prefrontal cortex by Western Blott 1 month after sham (white bars) or whole-brain irradiation with a single dose of 11 Gy (black bars). Data are expressed as means ± SEM. * p < 0.05 compared with the sham group using the Mann–Whitney U test

Article Snippet: Next, membranes were incubated overnight at 4 °C with either anti-GABAb mouse monoclonal antibody (1:1000, SC-166408, Santa Cruz, USA), anti-GABAa mouse monoclonal antibody (1:1000, sc-376282, Santa Cruz, USA), anti-NR1 monoclonal mouse antibody (1:200, BML-SA493-0015, Enzo, USA) or anti-NR2A monoclonal mouse antibody (1:200, sc-31540, Santa Cruz, USA) diluted in TBST containing 5% nonfat dry milk.

Techniques: Expressing, Western Blot, Irradiation, MANN-WHITNEY