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使用敲除细胞株进行验证重组RabMAb

重组Anti-NMDAR1抗体[EPR23397-66] - BSA and Azide free (ab274390)

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Western blot - Anti-NMDAR1 antibody [EPR23397-66] - BSA and Azide free (ab274390)
  • Western blot - Anti-NMDAR1 antibody [EPR23397-66] - BSA and Azide free (ab274390)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NMDAR1 antibody [EPR23397-66] - BSA and Azide free (ab274390)
  • Western blot - Anti-NMDAR1 antibody [EPR23397-66] - BSA and Azide free (ab274390)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NMDAR1 antibody [EPR23397-66] - BSA and Azide free (ab274390)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NMDAR1 antibody [EPR23397-66] - BSA and Azide free (ab274390)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NMDAR1 antibody [EPR23397-66] - BSA and Azide free (ab274390)
  • Anti-NMDAR1 antibody [EPR23397-66] - BSA and Azide free (ab274390)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23397-66] to NMDAR1 - BSA and Azide free
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 647 Unconjugated

概述

  • 产品名称

    Anti-NMDAR1抗体[EPR23397-66] - BSA and Azide free
    参阅全部 NMDAR1 一抗

  • 描述

    兔单克隆抗体[EPR23397-66] to NMDAR1 - BSA and Azide free

  • 宿主

    Rabbit

  • 经测试应用

    适用于: WB, IHC-Pmore details
    不适用于: Flow Cyt,ICC/IF,IHC-Fr or IP

  • 种属反应性

    与反应: Mouse, Rat, Human

  • 免疫原

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • 阳性对照

    • WB: Mouse brain tissue lysate; Rat brain tissue lysate; Human brain tissue lysate; Wild-type Neuro-2a cell lysate. IHC-P: Human cerebrum tissue; Mouse cerebrum tissue; rat cerebrum tissue.

  • 常规说明

    ab274390 is the carrier-free version of ab274377.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab274390于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
WB
Use at an assay dependent concentration. Detects a band of approximately 120 kDa (predicted molecular weight: 105 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
说明
WB
Use at an assay dependent concentration. Detects a band of approximately 120 kDa (predicted molecular weight: 105 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
应用说明
Is unsuitable for Flow Cyt,ICC/IF,IHC-Fr or IP.

靶标

  • 功能

    NMDA receptor subtype of glutamate-gated ion channels with high calcium permeability and voltage-dependent sensitivity to magnesium. Mediated by glycine. This protein plays a key role in synaptic plasticity, synaptogenesis, excitotoxicity, memory acquisition and learning. It mediates neuronal functions in glutamate neurotransmission. Is involved in the cell surface targeting of NMDA receptors.

  • 序列相似性

    Belongs to the glutamate-gated ion channel (TC 1.A.10.1) family. NR1/GRIN1 subfamily.

  • 翻译后修饰

    NMDA is probably regulated by C-terminal phosphorylation of an isoform of NR1 by PKC. Dephosphorylated on Ser-897 probably by protein phosphatase 2A (PPP2CB). Its phosphorylated state is influenced by the formation of the NMDAR-PPP2CB complex and the NMDAR channel activity.

  • 细胞定位

    Cell membrane. Cell junction > synapse > postsynaptic cell membrane. Cell junction > synapse > postsynaptic cell membrane > postsynaptic density. Enriched in post-synaptic plasma membrane and post-synaptic densities.
  • Information by UniProt

  • 数据库链接

    see all

  • 别名

    • GluN1 antibody
    • Glutamate [NMDA] receptor subunit zeta-1 antibody
    • Glutamate receptor ionotropic N methyl D aspartate 1 antibody
    • Glutamate receptor ionotropic, N-methyl-D aspartate, subunit 1 antibody
    • glutamate receptor ionotropic, NMDA 1 antibody
    • Grin1 antibody
    • MRD8 antibody
    • N methyl D aspartate receptor antibody
    • N methyl D aspartate receptor channel subunit zeta 1 antibody
    • N methyl D aspartate receptor subunit NR1 antibody
    • N-methyl-D-aspartate receptor subunit NR1 antibody
    • NMD-R1 antibody
    • NMDA 1 antibody
    • NMDA R1 antibody
    • NMDA receptor 1 antibody
    • NMDA1 antibody
    • NMDAR antibody
    • NMDZ1_HUMAN antibody
    • NR1 antibody
    see all

图片

  • Western blot - Anti-NMDAR1 antibody [EPR23397-66] - BSA and Azide free (ab274390)
    Western blot - Anti-NMDAR1 antibody [EPR23397-66] - BSA and Azide free (ab274390)
    All lanes : Anti-NMDAR1 antibody [EPR23397-66] (ab274377) at 1/1000 dilution

    Lane 1 : Wild-type Neuro-2a cell lysate
    Lane 2 : GRIN1 knockout Neuro-2a cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 105 kDa
    Observed band size: 105-125 kDa why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab274377).

    Anti-NMDAR1 antibody [EPR23397-66] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab274377 was shown to bind specifically to NMDAR1. A band was observed at 105-125 kDa in wild-type Neuro-2a cell lysates with no signal observed at this size in GRIN1 knockout cell line ab281960 (knockout cell lysate ab282987). To generate this image, wild-type and GRIN1 knockout Neuro-2a cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween®20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  • Western blot - Anti-NMDAR1 antibody [EPR23397-66] - BSA and Azide free (ab274390)
    Western blot - Anti-NMDAR1 antibody [EPR23397-66] - BSA and Azide free (ab274390)
    All lanes : Anti-NMDAR1 antibody [EPR23397-66] (ab274377) at 1/1000 dilution

    Lane 1 : Mouse brain tissue lysate
    Lane 2 : Mouse liver tissue lysate
    Lane 3 : Mouse heart tissue lysate
    Lane 4 : Rat brain tissue lysate
    Lane 5 : Rat liver tissue lysate
    Lane 6 : Rat heart tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

    Predicted band size: 105 kDa
    Observed band size: 120 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    The molecular weight observed is consistent with what has been described in the literature (PMID: 10818139).

    Negative control: liver and heart (PMID: 10480904).

    Samples are non-boiled as boiling may cause protein aggregates.

    Exposure time: 5.5 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab274377)

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NMDAR1 antibody [EPR23397-66] - BSA and Azide free (ab274390)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NMDAR1 antibody [EPR23397-66] - BSA and Azide free (ab274390)

    Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling NMDAR1 with abab274377 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on mouse cerebrum (PMID: 27391811). The section was incubated with ab274377 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab274377).

  • Western blot - Anti-NMDAR1 antibody [EPR23397-66] - BSA and Azide free (ab274390)
    Western blot - Anti-NMDAR1 antibody [EPR23397-66] - BSA and Azide free (ab274390)
    All lanes : Anti-NMDAR1 antibody [EPR23397-66] (ab274377) at 1/1000 dilution

    Lane 1 : Human brain tissue lysate
    Lane 2 : Human heart tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

    Predicted band size: 105 kDa
    Observed band size: 120 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    Samples are non-boiled as boiling may cause protein aggregates.

    Exposure time: 15 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab274377)

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NMDAR1 antibody [EPR23397-66] - BSA and Azide free (ab274390)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NMDAR1 antibody [EPR23397-66] - BSA and Azide free (ab274390)

    Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling NMDAR1 with abab274377 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on rat cerebrum (PMID: 27391811). The section was incubated with ab274377 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab274377).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NMDAR1 antibody [EPR23397-66] - BSA and Azide free (ab274390)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NMDAR1 antibody [EPR23397-66] - BSA and Azide free (ab274390)

    Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling NMDAR1 with abab274377 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control: no staining on human liver (PMID: 10480904). The section was incubated with ab274377 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab274377).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NMDAR1 antibody [EPR23397-66] - BSA and Azide free (ab274390)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NMDAR1 antibody [EPR23397-66] - BSA and Azide free (ab274390)

    Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling NMDAR1 with ab274377 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human cerebrum (PMID: 27391811). The section was incubated with ab274377 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab274377).

  • Anti-NMDAR1 antibody [EPR23397-66] - BSA and Azide free (ab274390)
    Anti-NMDAR1 antibody [EPR23397-66] - BSA and Azide free (ab274390)

实验方案

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

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