NMDAR2A Antibody - C-terminus - Azide Free

Novus Biologicals | Catalog # NB300-105

Novus Biologicals
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Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Rabbit

Cited:

Human, Mouse, Rat

Applications

Validated:

Knockout Validated, Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunoprecipitation

Cited:

Western Blot, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, IF/IHC

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

Azide Free
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Product Specifications

Immunogen

Fusion protein from the C-terminal region of the NR2A subunit. Accession # Q00959

Reactivity Notes

Use in Mouse reported in scientific literature (PMID:35354047).

Localization

NB 300-105 can be used to demonstrate that both the NR2A is highly enriched in hippocampus compared to either cortex or cerebellum.

Specificity

Specific for endogenous levels of the ~180 kDa NR2A subunit of the NMDA receptor. No reactivity towards the NR2B and NR2C subunits. Immunolabeling is blocked by pre-adsorption of antibody with the fusion protein used to generate the antibody.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

180 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Description

PLEASE NOTE: If 0.01mg size is ordered, product will come lyophilized from 5 mM ammonium bicarbonate. Please reconstitute lyophilized product in 50 ul phosphate buffered saline (PBS: 137 mM NaCl, 7.5 mM Na2HPO4, 2.7 mM KCl, 1.5 mM KH2PO4, pH 7.4)

Scientific Data Images for NMDAR2A Antibody - C-terminus - Azide Free

Western Blot: NMDAR2A Antibody [NB300-105]

Western Blot: NMDAR2A Antibody [NB300-105]

NMDAR2A-Antibody-Western-Blot-NB300-105-img0006.jpg
Western Blot: NMDAR2A Antibody [NB300-105]

Western Blot: NMDAR2A Antibody [NB300-105]

NMDAR2A-Antibody-Western-Blot-NB300-105-img0007.jpg
Immunohistochemistry: NMDAR2A Antibody [NB300-105]

Immunohistochemistry: NMDAR2A Antibody [NB300-105]

Immunohistochemistry: NMDAR2A Antibody [NB300-105] - Immunostaining of rabbit retina showing NR2A in the rod and cone photoreceptors in the outer plexiform layer as well as the entire inner plexiform layer.
Western Blot: NMDAR2A Antibody [NB300-105]

Western Blot: NMDAR2A Antibody [NB300-105]

Western Blot: NMDAR2A Antibody [NB300-105]
NMDAR2A Antibody

Western Blot: NMDAR2A Antibody [NB300-105] -

Western Blot: NMDAR2A Antibody [NB300-105] - NMDAR subunit composition is immature in whole lysates from Ppt1-/- visual cortex.(A) Representative immunoblots of the GluN2A in whole lysates across age & genotype as indicated (top) & quantification of band density (bottom) normalized to beta -actin loading control within lane. (B) Representative immunoblots of the GluN2B in whole lysates across age & genotype as indicated (top) & quantification of band density (bottom) normalized to beta -actin loading control within lane. (C) Representative immunoblots of GluN2A & GluN2B (top) from whole lysates across age & genotype & quantification of the ratio of GluN2A/GluN2B band density within animal (bottom). (D) Representative immunoblots of GluN1 in whole lysates across age & genotype as indicated (top) & quantification of band density (bottom) normalized to beta -actin loading control within lane. (E) Representative immunoblot from whole lysates of PPT1 across age & genotype as indicated (top) & protein expression level (bottom) normalized to beta -actin. For experiments in Figure 2—figure supplement 1A–C, Ppt1-/- & WT were compared (n = 4 independent experiments/animals with two repetitions/group) at each age using t-test & the significance is indicated: *p<0.05. In Figure 2D, WT expression levels at each age were compared (n = 4 independent experiments/animals with two repetitions/group) by ANOVA followed by Tukey’s post-hoc test. Significance between ages is indicated: *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. Error bars represent s.e.m. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30946007), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
NMDAR2A Antibody

Western Blot: NMDAR2A Antibody [NB300-105] -

Western Blot: NMDAR2A Antibody [NB300-105] - GluN2B to GluN2A NMDAR switch & Ppt1-/--induced synaptic deficits are recapitulated in primary cortical neurons.(A) Representative immunoblot (top) & quantification of GluN2B levels in WT & Ppt1-/- neurons at DIV7, 10, & 18. (B) Representative immunoblot (top) & quantification of GluN2A levels (bottom) in WT & Ppt1-/- neurons at DIV7, 10, & 18. (C) Representative immunoblot (top) & quantification of PSD-95 levels (bottom) in WT & Ppt1-/- neurons at DIV7, 10, & 18. (D) Representative immunoblot (top) & quantification of the GluN2A/2B ratio (bottom) in WT & Ppt1-/- neurons at DIV7, 10, & 18. For all experiments in Figure 5, Ppt1-/- & WT were compared (n = 2 independent experiments with two repetitions/group) at each time point using t-test & the significance indicated as follows: *p<0.05 where indicated. Error bars represent s.e.m. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30946007), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
NMDAR2A Antibody

Western Blot: NMDAR2A Antibody [NB300-105] -

Western Blot: NMDAR2A Antibody [NB300-105] - NMDAR subunit composition is immature in whole lysates from Ppt1-/- visual cortex.(A) Representative immunoblots of the GluN2A in whole lysates across age & genotype as indicated (top) & quantification of band density (bottom) normalized to beta -actin loading control within lane. (B) Representative immunoblots of the GluN2B in whole lysates across age & genotype as indicated (top) & quantification of band density (bottom) normalized to beta -actin loading control within lane. (C) Representative immunoblots of GluN2A & GluN2B (top) from whole lysates across age & genotype & quantification of the ratio of GluN2A/GluN2B band density within animal (bottom). (D) Representative immunoblots of GluN1 in whole lysates across age & genotype as indicated (top) & quantification of band density (bottom) normalized to beta -actin loading control within lane. (E) Representative immunoblot from whole lysates of PPT1 across age & genotype as indicated (top) & protein expression level (bottom) normalized to beta -actin. For experiments in Figure 2—figure supplement 1A–C, Ppt1-/- & WT were compared (n = 4 independent experiments/animals with two repetitions/group) at each age using t-test & the significance is indicated: *p<0.05. In Figure 2D, WT expression levels at each age were compared (n = 4 independent experiments/animals with two repetitions/group) by ANOVA followed by Tukey’s post-hoc test. Significance between ages is indicated: *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. Error bars represent s.e.m. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30946007), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
NMDAR2A Antibody

Western Blot: NMDAR2A Antibody [NB300-105] -

Western Blot: NMDAR2A Antibody [NB300-105] - GluN2B to GluN2A NMDAR switch & Ppt1-/--induced synaptic deficits are recapitulated in primary cortical neurons.(A) Representative immunoblot (top) & quantification of GluN2B levels in WT & Ppt1-/- neurons at DIV7, 10, & 18. (B) Representative immunoblot (top) & quantification of GluN2A levels (bottom) in WT & Ppt1-/- neurons at DIV7, 10, & 18. (C) Representative immunoblot (top) & quantification of PSD-95 levels (bottom) in WT & Ppt1-/- neurons at DIV7, 10, & 18. (D) Representative immunoblot (top) & quantification of the GluN2A/2B ratio (bottom) in WT & Ppt1-/- neurons at DIV7, 10, & 18. For all experiments in Figure 5, Ppt1-/- & WT were compared (n = 2 independent experiments with two repetitions/group) at each time point using t-test & the significance indicated as follows: *p<0.05 where indicated. Error bars represent s.e.m. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30946007), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for NMDAR2A Antibody - C-terminus - Azide Free

Application
Recommended Usage

Immunohistochemistry

1:1000 - 1:2000

Immunohistochemistry-Paraffin

1:1000

Immunoprecipitation

3 ul per 200 ug lysate

Western Blot

1:1000
Application Notes
NB 300-105 can be used in Western blot where a band is seen at ~ 180 kDa representing the NMDAR2A.

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Formulation

10mM HEPES (pH 7.5), 0.15M NaCl, 0.1 mg/ml BSA and 50% Glycerol

Format

Azide Free

Preservative

No Preservative

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C. Avoid freeze-thaw cycles.

Background: NMDAR2A

The ion channels activated by glutamate are divided into two classes. Those that are sensitive to N-methyl-D-aspartate (NMDA) are designated NMDA receptors (NMDAR) while those activated by kainate and a-amino-3-hydroxy-5-methyl-4-isoxalone propionic acid (AMPA) are known as kainate/AMPA receptors (K/AMPAR). NMDA receptors are among the most studied receptors in neuroscience because they are involved in neuronal cell development and plasticity, a cellular correlate for learning. NMDA receptors are also implicated in several disorders of the central nervous system including epilepsy and ischemic neuronal cell death. NMDA receptors also appear to be a target for ethanol at physiological concentrations and therefore may play a significant role in alcoholism.

Alternate Names

GluN2A, glutamate receptor, ionotropic, N-methyl D-aspartate 2A, hNR2A, NMDA receptor subtype 2A, N-methyl D-aspartate receptor subtype 2A, N-methyl-D-aspartate receptor subunit 2A, subunit epsilon-1

Entrez Gene IDs

2903 (Human); 24409 (Rat)

Gene Symbol

GRIN2A

UniProt

Additional NMDAR2A Products

Product Documents for NMDAR2A Antibody - C-terminus - Azide Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Product Specific Notices for NMDAR2A Antibody - C-terminus - Azide Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for NMDAR2A Antibody - C-terminus - Azide Free

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Protocols

View specific protocols for NMDAR2A Antibody - C-terminus - Azide Free (NB300-105):

Western Blot Protocol specific for NMDAR2A Antibody (NB300-105):
Western Blot:

**For brain tissue homogenize in hot 1% SDS then run 7.5 % (as 2A is ~ 180 kDa) standard SDS gels and blot.

1. Thoroughly sonicate cell lysates or tissue homogenates to be loaded onto gel then dilute in appropriate sample buffer and boil for 5 minutes at 100C. Let samples cool to room temperature then load onto gel.
2. Run SDS-PAGE per gel apparatus manufacturer'??s instructions.
3. Transfer proteins to nitrocellulose or PVDF membrane (if using PVDF, be sure to activate membrane in Methanol prior to use).
4. After transfer, air-dry blot to more stably fix proteins onto membrane.
5. Block non-specific sites on membrane in 5% NFDM (Non-fat dry milk) or 3% BSA-TTBS (Tris-Buffered Saline + 0.1% Tween-20) for 1 hour while shaking at room temperature.
6. Incubate membrane in primary antibody diluted in 1% NFDM (or BSA)-TTBS while shaking overnight at 4C.
7. Decant unbound primary antibody solution and wash blot 3 x 10 minutes in TTBS.
8. Incubate blots in appropriate HRP-conjugated (for ECL detection) secondary antibody at a 1:10,000 -1:20,000 dilution in 1% Milk (or BSA)-TTBS for 1 hour while shaking at room temperature.
9. Decant secondary antibody solution and wash blots 3 x 10 minutes in TTBS or use TBS + 0.1% Triton(R) X-100 to reduce excessive background if needed.
10. ECL Detect.

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FAQs

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