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Purified Mouse Anti-NMDAR1
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Purified Mouse Anti-NMDAR1
Western blot analysis of NMDAR1 (left panel). Lysate from a rat cortex was probed with Purified Mouse Anti-NMDAR1 (Cat. No. 556308) at concentrations of 3 µg/mL (lane 1), 1.0 µg/mL (lane 2), and 0.5 µg/ml (lane 3). NMDAR1 can be recognized as a band of ~120 kDa. Immunocytochemical detection of NMDAR1 (center panels). HEK-293 cells (Human embryonic kidney cells; ATCC CRL-1573) transfected with NMDAR1 (panel B) or untransfected (panel C) were stained with Purified Mouse Anti-NMDAR1antibody. Arrows in the left panel indicate examples of NMDAR1 immunoreactive cells. Immunofluorescent staining of differentiated SH-SY5Y cells (Human neuroblastoma; ATCC CRL-2266) (right panel). Cells were seeded in a 96-well, collagen coated imaging plate at ~5,000 cells per well.  Cells were incubated with 50 µM ATRA (all-trans-Retinoic-acid) (Sigma-Aldrich; cat.no. R2625) for 5 days, followed by 50 ng/ml BDNF (Sigma-Aldrich; cat.no. B3795)  for 5 days.  Differentiated cells were fixed and stained using the methanol fix/perm protocol (see Recommended Assay Procedure) and the Purified Mouse Anti-NMDAR1 antibody.  The second step reagent was Alexa Fluor® 488 goat anti-mouse Ig (Invitrogen).  The image was taken on a BD Pathway™ 855 Bioimager using a 20x objective.  This antibody also stained undifferentiated SH-SY5Y, SK-N-SH (Human neuroblastoma; ATCC HTB-11), and C6 cells (Rat glioma; ATCC CCL-107) using both the Triton-X 100 and methanol fix/perm protocols (see Recommended Assay Procedure).
Western blot analysis of NMDAR1 (left panel). Lysate from a rat cortex was probed with Purified Mouse Anti-NMDAR1 (Cat. No. 556308) at concentrations of 3 µg/mL (lane 1), 1.0 µg/mL (lane 2), and 0.5 µg/ml (lane 3). NMDAR1 can be recognized as a band of ~120 kDa. Immunocytochemical detection of NMDAR1 (center panels). HEK-293 cells (Human embryonic kidney cells; ATCC CRL-1573) transfected with NMDAR1 (panel B) or untransfected (panel C) were stained with Purified Mouse Anti-NMDAR1antibody. Arrows in the left panel indicate examples of NMDAR1 immunoreactive cells. Immunofluorescent staining of differentiated SH-SY5Y cells (Human neuroblastoma; ATCC CRL-2266) (right panel). Cells were seeded in a 96-well, collagen coated imaging plate at ~5,000 cells per well.  Cells were incubated with 50 µM ATRA (all-trans-Retinoic-acid) (Sigma-Aldrich; cat.no. R2625) for 5 days, followed by 50 ng/ml BDNF (Sigma-Aldrich; cat.no. B3795)  for 5 days.  Differentiated cells were fixed and stained using the methanol fix/perm protocol (see Recommended Assay Procedure) and the Purified Mouse Anti-NMDAR1 antibody.  The second step reagent was Alexa Fluor® 488 goat anti-mouse Ig (Invitrogen).  The image was taken on a BD Pathway™ 855 Bioimager using a 20x objective.  This antibody also stained undifferentiated SH-SY5Y, SK-N-SH (Human neuroblastoma; ATCC HTB-11), and C6 cells (Rat glioma; ATCC CCL-107) using both the Triton-X 100 and methanol fix/perm protocols (see Recommended Assay Procedure).
Product Details
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BD Pharmingen™
N-Methyl-D-Aspartate Receptor-1; Glutamate Receptor
Rat (QC Testing), Human, Monkey (Reported)
Mouse IgG2a
Rat NMDAR1 aa. 660-811 Recombinant Protein
Western blot (Routinely Tested), Bioimaging, Immunohistochemistry (Tested During Development), Electron microscopy (Reported)
120 kDa
0.5 mg/ml
AB_396353
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Recommended Assay Procedures

Western blot:  Please refer to "Cell Biology (WB, IP, IHC, IF)" on our website at: http://www.bdbiosciences.com/us/s/resources

Immunohistochemistry: Investigators are encouraged to titrate the antibody, such as a range including a 1:250-1:1000 dilution.

Bioimaging:

Methanol Procedure for a 96 well plate:

Remove media from wells. Add 100 µl/well fresh 3.7% Formaldehyde in PBS. Incubate for 10 minutes at room temperature (RT). Flick out and add 100 µl/well 90% methanol. Incubate for 5 minutes at RT. Flick out and wash twice with PBS. Flick out PBS and add 100 µl/well blocking buffer (3% FBS in PBS). Incubate for 30 minutes at RT. Flick out and add diluted antibody (diluted in blocking buffer). Incubate for 1 hour at RT. Wash three times with PBS. Flick out PBS and add second step reagent. Incubate for 1 hour at RT. Wash three times with PBS. Image sample.

Triton-X 100 Procedure for a 96 well plate:

Remove media from wells. Add 100 µl/well fresh 3.7% Formaldehyde in PBS. Incubate for 10 minutes at room temperature (RT). Flick out and add 100 µl/well 0.1% Triton-X 100. Incubate for 5 minutes at RT. Flick out and wash twice with PBS. Flick out PBS and add 100 µl/well blocking buffer (3% FBS in PBS). Incubate for 30 minutes at RT. Flick out and add diluted antibody (diluted in blocking buffer). Incubate for 1 hour at RT. Flick out and wash three times with PBS. Flick out and add second step reagent. Incubate for 1 hour at RT. Flick out and wash three times with PBS. Image sample.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  3. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  4. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
556308 Rev. 13
Antibody Details
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54.1

Glutamate is a major excitatory neurotransmitter in mammalian brain. Glutamatergic neurotransmission is mediated by a family of glutamate receptors. They can be grouped into two classes, ionotropic (GluR) and metabotropic (mGluR) receptors. The ionotropic GluRs can be divided into two subclasses, N-Methyl-D-Aspartate (NMDA) and non-NMDA receptors, based on physiological studies and affinity to different glutamate agonists. At least six different forms of NMDA receptors have been reported, NMDA -R1, -R2, -R2A, -R2B, -R2C, and -R2D. NMDAR1 has been reported to be required for the formation of functional NMDA receptors and can be observed migrating at ~120 kDa in SDS/PAGE. This antibody has been reported not to cross-react with NMDAR2-5 receptors.

556308 Rev. 13
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
556308 Rev.13
Citations & References
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View product citations for antibody "556308" on CiteAb

Development References (4)

  1. Brose N, Huntley GW, Stern-Bach Y, Sharma G, Morrison JH, Heinemann SF. Differential assembly of coexpressed glutamate receptor subunits in neurons of rat cerebral cortex. J Biol Chem. 1994; 269(24):16780-16784. (Biology: Electron microscopy, Western blot). View Reference
  2. Siegel SJ, Brose N, Janssen WG, et al. Regional, cellular, and ultrastructural distribution of N-methyl-D-aspartate receptor subunit 1 in monkey hippocampus. Proc Natl Acad Sci U S A. 1994; 91(2):564-568. (Immunogen: Electron microscopy, Western blot). View Reference
  3. Siegel SJ, Janssen WG, Tullai JW, et al. Distribution of the excitatory amino acid receptor subunits GluR2(4) in monkey hippocampus and colocalization with subunits GluR5-7 and NMDAR1. J Neurosci. 1995; 15(4):2707-2719. (Biology: Electron microscopy). View Reference
  4. Wood MW, VanDongen HM, VanDongen AM. Structural conservation of ion conduction pathways in K channels and glutamate receptors. Proc Natl Acad Sci U S A. 1995; 92(11):4882-4886. (Biology: Western blot). View Reference
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556308 Rev. 13

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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.