Anti-NMDAR2B antibody

5 product images

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  Western blot - Anti-NMDAR2B antibody (ab65783)

  NMDAR2B contains a number of potential phosphorylation and glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.

  All lanes: Western blot - Anti-NMDAR2B antibody (ab65783) at 1 µg/mL

  Lane 1: Human brain tissue lysate - total protein (ab29466) at 10 µg

  Lane 2: Brain (Mouse) Tissue Lysate at 10 µg

  Lane 3: Brain (Rat) Tissue Lysate at 10 µg

  Lane 4: Hippocampus (Mouse) Tissue Lysate at 10 µg

  Lane 5: Rat Hippocampus Tissue Lysate at 10 µg

  Secondary

  All lanes: Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

  Performed under reducing conditions.

  Predicted band size: 166 kDa

  Observed band size: 180 kDa, 35 kDa

  Exposure time: 1min

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  Immunoprecipitation - Anti-NMDAR2B antibody (ab65783)

  NMDAR2B was immunoprecipitated using 0.5mg Mouse Brain tissue lysate, 5µg of Rabbit polyclonal to NMDAR2B and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
  The antibody was incubated under agitation with Protein G beads for 10min, Mouse Brain tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
  Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70^oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab65783.
  Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) ab99697).
  Band: 180kDa; NMDAR2B

  All lanes: Immunoprecipitation - Anti-NMDAR2B antibody (ab65783)

  Predicted band size: 166 kDa

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  Western blot - Anti-NMDAR2B antibody (ab65783)

  This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab65783 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ECL Substrate Kit (High Sensitivity) ab133406.

  All lanes: Western blot - Anti-NMDAR2B antibody (ab65783) at 1 µg/mL

  Lane 1: Rat Hippocampus Tissue Lysate at 10 µg

  Lane 2: Mouse Hippocampus Tissue Lysate at 10 µg

  Lane 3: Human brain tissue lysate - total protein (ab29466) at 20 µg

  Lane 4: Human brain hippocampus tissue lysate - total protein (ab30180) at 20 µg

  Lane 5: Human brain amygdala tissue lysate - total protein at 10 µg

  Secondary

  All lanes: Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 166 kDa

  Observed band size: 100 kDa, 180 kDa, 200 kDa, 35 kDa, 45 kDa, 56 kDa, 65 kDa

  Exposure time: 8min

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  Immunocytochemistry/ Immunofluorescence - Anti-NMDAR2B antibody (ab65783)

  ab65783 staining NMDAR2B in Rat Primary Neurons DIV14 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab65783 at 1µg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).

  Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown..

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  Image collected and cropped by CiteAb under a CC-BY license from the publication

  Western blot - Anti-NMDAR2B antibody (ab65783)

  NMDAR2B western blot using anti-NMDAR2B antibody ab65783. Publication image and figure legend from Colas, J., Chessel, N., et al., 2020, Neural Plast, PubMed 32256561.

  
  

  ab65783 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab65783 please see the product overview.

  Changes in protein expression following 1 μM all-trans-retinoic acid (RA) treatment of the deep and superficial neocortical layers, as well as of the hippocampus, of (a) three 1-month-old and (b) three 17-month-old male C57BL/6J mice. The proteins are involved in the amyloid cascade (Presenilin 1 or PS1/γ-secretase, BACE1/β-secretase, ADAM10/α-secretase, and APP C-terminus), in Tau phosphorylation (phospho-Tau (AT8) or unphosphorylated Tau (Tau1)), in synaptic functions (PSD95, GluN2B/NR2B), or in RA-dependent pathways (RARα, PPARβ/δ). GAPDH and DB71 stainings were used to demonstrate equal loading of the Western blot gels. Overall, we observed significant increases (see Section 3.3) of ADAM10, APP, and phosphorylated and unphosphorylated Tau proteins, suggesting the activation of neuroprotective mechanisms following the RA treatment, whereas the expression of the enzymes of the amyloidogenic pathway, PS1 and BACE1, or, in most cases, of the RA receptors was not increased. Protein sizes are indicated on the right. A size range is given for Tau isoforms (AT8 and Tau1).