Anti-NMDAR2B antibody [NR2B] (ab28373)

Reviews (3)Q&A (3)References (6)

Overview

  • Product name
    Anti-NMDAR2B antibody [NR2B]
    See all NMDAR2B primary antibodies
  • Description
    Mouse monoclonal [NR2B] to NMDAR2B
  • Host species
    Mouse
  • Specificity
    Detects the N-methyl-D-aspartate (NMDA) receptor type 2B. This antibody has been tested in mouse and rat brain in western blotting. It has not been tested in human for Western blotting - only for ICC on transfected HEK cells.
  • Tested applications
    Suitable for: ICC, WB, IHC-P, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment corresponding to Rat NMDAR2B aa 934-1457.

  • Positive control
    • WB: Mouse brain tissue ICC: transfected HEK cells

Properties

Applications

Our Abpromise guarantee covers the use of ab28373 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC Use a concentration of 1 µg/ml.
WB Use a concentration of 2 µg/ml. Predicted molecular weight: 166 kDa.
IHC-P 1/10 - 1/100.
Flow Cyt Use 1-2µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

Target

  • Function
    NMDA receptor subtype of glutamate-gated ion channels with high calcium permeability and voltage-dependent sensitivity to magnesium. Mediated by glycine.
  • Tissue specificity
    Primarily found in the fronto-parieto-temporal cortex and hippocampus pyramidal cells, lower expression in the basal ganglia.
  • Sequence similarities
    Belongs to the glutamate-gated ion channel (TC 1.A.10.1) family. NR2B/GRIN2B subfamily.
  • Cellular localization
    Cell membrane. Cell junction > synapse > postsynaptic cell membrane.
  • Information by UniProt
  • Database links
    see all
  • Alternative names
    • AW490526 antibody
    • EIEE27 antibody
    • Glutamate [NMDA] receptor subunit epsilon 2 antibody
    • Glutamate [NMDA] receptor subunit epsilon-2 antibody
    • Glutamate Receptor Ionotropic N Methyl D Aspartate 2B antibody
    • Glutamate Receptor Ionotropic N Methyl D Aspartate subunit 2B antibody
    • Glutamate receptor ionotropic NMDA2B antibody
    • Glutamate receptor subunit epsilon 2 antibody
    • Glutamate receptor, ionotropic, NMDA2B (epsilon 2) antibody
    • GRIN 2B antibody
    • GRIN2B antibody
    • hNR 3 antibody
    • hNR3 antibody
    • MGC142178 antibody
    • MGC142180 antibody
    • MRD6 antibody
    • N methyl D asparate receptor channel subunit epsilon 2 antibody
    • N methyl D aspartate receptor subtype 2B antibody
    • N methyl D aspartate receptor subunit 2B antibody
    • N methyl D aspartate receptor subunit 3 antibody
    • N-methyl D-aspartate receptor subtype 2B antibody
    • N-methyl-D-aspartate receptor subunit 3 antibody
    • NMDA NR2B antibody
    • NMDA R2B antibody
    • NMDAR2B antibody
    • NMDE2 antibody
    • NMDE2_HUMAN antibody
    • NME2 antibody
    • NR2B antibody
    • NR3 antibody
    see all

Images

  • Western blot - Anti-NMDAR2B antibody [NR2B] (ab28373)
    Western blot - Anti-NMDAR2B antibody [NR2B] (ab28373)
    Anti-NMDAR2B antibody [NR2B] (ab28373) at 2 µg/ml + mouse brain tissue

    Predicted band size: 166 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NMDAR2B antibody [NR2B] (ab28373)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NMDAR2B antibody [NR2B] (ab28373)

    ab28373 staining NMDAR2B in Human breast tissue sections (right) compared with negative control (left) by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 3% H2O2-methanol for 15 minutes at room temperature; antigen retrieval was by heat mediation in a sodium citrate buffer. Samples were incubated with primary antibody (1/20) overnight at 4°C. A HRP-conjugated secondary antibody was used for detection.

  • Western blot - Anti-NMDAR2B antibody [NR2B] (ab28373)
    Western blot - Anti-NMDAR2B antibody [NR2B] (ab28373)
    Lanes 2-4 : Anti-NMDAR2B antibody [NR2B] (ab28373) at 1/500 dilution
    Lanes 5-7 : Anti-NMDAR2B antibody [NR2B] (ab28373) at 1/1000 dilution

    Lanes 3 & 6 : Mouse brain (50ug)
    Lanes 4 & 7 : Rat brain (50ug)

    Predicted band size: 166 kDa

  • Western blot - Anti-NMDAR2B antibody [NR2B] (ab28373)
    Western blot - Anti-NMDAR2B antibody [NR2B] (ab28373)This image is courtesy of an anonymous Abreview
    Anti-NMDAR2B antibody [NR2B] (ab28373) at 1/500 dilution + Rat P14 Cortex Lysate, vesicular fraction at 5 µg

    Secondary
    HRP conjugated Donkey anti-mouse IgG at 1/15000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 166 kDa



    Samples were incubated with primary antibody for 12 hours at 4ºC.

    Blocked with 5% milk for 1 hour at 22ºC. 

    See Abreview

  • Flow Cytometry - Anti-NMDAR2B antibody [NR2B] (ab28373)
    Flow Cytometry - Anti-NMDAR2B antibody [NR2B] (ab28373)
    Overlay histogram showing SH-SY5Y cells stained with ab28373 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab28373, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References

This product has been referenced in:
  • Zhou D  et al. GLYX-13 Ameliorates Schizophrenia-Like Phenotype Induced by MK-801 in Mice: Role of Hippocampal NR2B and DISC1. Front Mol Neurosci 11:121 (2018). Read more (PubMed: 29695955) »
  • Posa L  et al. Methadone Reverses Analgesic Tolerance Induced by Morphine Pretreatment. Int J Neuropsychopharmacol 19:N/A (2016). WB ; Mouse . Read more (PubMed: 26390873) »
See all 6 Publications for this product

Publishing research using ab28373? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-6 of 6 Abreviews or Q&A

Western blot abreview for Anti-NMDAR2B antibody [NR2B]

 Poor
Abreviews
abreview image
Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (stacking gel: 4%, resolving gel: 5-13%)
Sample
Rat Tissue lysate - other (hippocampi total membrane fraction)
Specification
hippocampi total membrane fraction
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Feb 19 2014

Question

Dear Abcam manager


I just send you the mail again to ask about the 2nd antibody for fluoroscence WB detection.

I sent you the specification information on chemidoc system that we want to use last week but didn't get reply yet.

According to BioRad web sites, each color emission filter can detect below dyes.



RED: Cy5, Cy5.5, Alexa Fluor 647 / 680, DyLight 649 / 680, IRDye 680

GREEN: Cy3, Flamingo, Krypton, Pro-Q Diamond, Alexa Fluor 546, DyLight 549, Rhodamine

BLUE: Cy2, Coomassie Fluor Orange, Alexa Fluor 488, DyLight 488, Pro-! Emerald, Qdot 605 / 625 / 705

Read More
Answer

Thank you for your message and for your patience.
It is difficult to find three secondary antibodies in three colors that woudl be suitable, but I hope the following will help. Please check the excitation and emission wavelenghts and spectra to ensure they are suitable for the machine you are using. (I say this because some of the blue dyes they suggest go into the green range in the light spectra). I can recommend to review the online datasheets for further information.
Anti NMDAR1 antibody (ab77264, goat polyclonal)
Red fluorophore Secondary antibodies for detection of goat primary:
Donkey anti-Goat IgG H&L (PE) secondary antibody (ab7004)
Donkey F(ab')2 polyclonal Secondary Antibody to Goat IgG - H&L (PE), pre-adsorbed
https://www.abcam.com/index.html?datasheet=7004 https://www.abcam.com/index.html?datasheet=7004.
Anti-NMDAR2A antibody (ab14596, rabbit polyclonal)
Green fluorophore Secondary antibodies for detection of rabbit primary:
ab6798 Donkey anti-Rabbit IgG H&L (FITC) secondary antibody
https://www.abcam.com/index.html?datasheet=6798 https://www.abcam.com/index.html?datasheet=6798.

ab7079 Donkey anti-Rabbit IgG H&L (FITC) secondary antibody
https://www.abcam.com/index.html?datasheet=7079 https://www.abcam.com/index.html?datasheet=7079.
Anti-NMDAR2B antibody (ab28373, mouse monoclonal)
The following may fit into the blue channel on the instrument you are using, but I can suggest to please check this. The emission spectra is slightly into the green spectra.
ab96875 Donkey anti-Mouse IgG H&L (DyLight® 488) secondary antibody
https://www.abcam.com/index.html?datasheet=96875 https://www.abcam.com/index.html?datasheet=96875.
I am sorry we do not have any that are tested and covered by the 6 month guarantee in western blot to suggest on this occasion.
The other option is to directly conjugate the primary antibodies yourself with our Easylink kits. These are quick and easy to use. However, please note that we recommend the following for these kits:
The antibody to be conjugated should be in 10-50mM amine-free buffer within a pH range of 6.5-8.5. Modest concentrations of Tris buffer (up to 20mM) are also tolerated.
I can recommend to read the following important information and frequently asked questions about the kits before selecting a suitable antibody to conjugate:
https://www.abcam.com/index.html?pageconfig=resource&rid=13148
https://www.abcam.com/index.html?pageconfig=resource&rid=13156
I hope this will be helpful. Please do not hesitate to contact me if you have any further questions.

Read More

Question

Currently using ab28373 in hamster tissue (IHC-Fr Fl), but not getting any signal.
Is there any NR2B antibodies that would work with hamster tissue.

Read More
Answer

Thank you for calling Abcam earlier today.

I have continued to try and find the hamster sequence for NR2B to see if there is any antibodies in our catalogue that may work for hamster, but I have not been able to find the hamster sequence.

There is a chance that ab65783 might work for you, but without the hamster sequence, I would feel unsure recommending it to you. If there is any chance that you could find the sequence and forward that to me, I would gladly do the alignments and see if there is a chance that it would work for you.

Please let me know if there is anything else I can help you with.

Read More

Question

Would like to try this antibody with hamster samples.

Read More
Answer

Thank you for your call today. I've issued the discount code below for testing ab28373in hamster samples.

DISCOUNT CODE: ***
Expiration date: July 30, 2012

This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for ab28373 in hamster samples and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Immunocytochemistry abreview for Anti-NMDAR2B antibody [NR2B]

 Average
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
Application
Immunocytochemistry
Sample
Mouse Cultured Cells (Hippocampal neurons)
Specification
Hippocampal neurons
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.25% Triton in PBS
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 21°C
Read More

Abcam user community

Verified customer

Submitted Dec 23 2011

Western blot abreview for Anti-NMDAR2B antibody [NR2B]

 Good
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
Application
Western blot
Sample
Rat Tissue lysate - whole (Rat P14 Cortex Lysate, vesicular fraction)
Loading amount
5 µg
Specification
Rat P14 Cortex Lysate, vesicular fraction
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More

Abcam user community

Verified customer

Submitted Dec 06 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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