NMDAR2A Antibody - C-terminus - Azide Free
Novus Biologicals | Catalog # NB300-105
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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.
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]
NMDAR2A-Antibody-Western-Blot-NB300-105-img0006.jpgImmunohistochemistry: 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] -
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.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.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.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
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
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.
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.
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Antigen Retrieval Protocol (PIER)
- Antigen Retrieval for Frozen Sections Protocol
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- Chromogenic IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Chromogenic Immunohistochemistry Staining of Frozen Tissue
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- Fluorescent IHC Staining of Frozen Tissue Protocol
- Graphic Protocol for Heat-induced Epitope Retrieval
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Graphic Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- IHC Sample Preparation (Frozen sections vs Paraffin)
- Immunofluorescent IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Immunohistochemistry Frozen Troubleshooting
- Immunohistochemistry Paraffin Troubleshooting
- Immunoprecipitation Protocol
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for Heat-Induced Epitope Retrieval (HIER)
- Protocol for Making a 4% Formaldehyde Solution in PBS
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Preparation & Fixation of Cells on Coverslips
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections - Graphic
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections - Graphic
- Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Immunohistochemistry
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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