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リコンビナントRabMAb

Anti-NP-I 抗体 [EPR25683-64] (ab289966)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NP-I antibody [EPR25683-64] (ab289966)
  • Immunoprecipitation - Anti-NP-I antibody [EPR25683-64] (ab289966)
  • Flow Cytometry (Intracellular) - Anti-NP-I antibody [EPR25683-64] (ab289966)
  • Western blot - Anti-NP-I antibody [EPR25683-64] (ab289966)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NP-I antibody [EPR25683-64] (ab289966)
  • Immunoprecipitation - Anti-NP-I antibody [EPR25683-64] (ab289966)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NP-I antibody [EPR25683-64] (ab289966)
  • Western blot - Anti-NP-I antibody [EPR25683-64] (ab289966)
  • Western blot - Anti-NP-I antibody [EPR25683-64] (ab289966)
  • Western blot - Anti-NP-I antibody [EPR25683-64] (ab289966)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NP-I antibody [EPR25683-64] (ab289966)
  • Anti-NP-I antibody [EPR25683-64] (ab289966)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR25683-64] to NP-I
  • Suitable for: IHC-P, IP, WB, Flow Cyt (Intra)
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

製品の概要

  • 製品名

    Anti-NP-I antibody [EPR25683-64]
    NP-I 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [EPR25683-64] to NP-I

  • 由来種

    Rabbit

  • 特異性

    This antibody does not react with Rat species for FC

  • アプリケーション

    適用あり: IHC-P, IP, WB, Flow Cyt (Intra)more details
    適用なし: ICC/IF

  • 種交差性

    交差種: Mouse, Rat, Human

  • 免疫原

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

  • ポジティブ･コントロール

    • WB: Human hypothalamus tissue lysate; Mouse hypothalamus tissue lysate; Rat brain and striatum tissue lysates; HeLa whole cell lysate. IHC-P: Human cerebrum tissue; Mouse cerebrum tissue; Rat cerebrum tissue. Flow Cyt (intra): HeLa cells. IP: HeLa whole cell lysate; Mouse hypothalamus tissue lysate.

  • 特記事項

    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.

製品の特性

  • 製品の状態

    Liquid

  • 保存方法

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.

  • バッファー

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...

  • 精製度

    Protein A purified

  • ポリ/モノ

    モノクローナル

  • クローン名

    EPR25683-64

  • アイソタイプ

    IgG

  • 研究分野

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab289966の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
1/30.
WB
1/1000. Detects a band of approximately 38, 50 kDa (predicted molecular weight: 47 kDa).
Flow Cyt (Intra)
1/500.

Not suitable with Rat.
特記事項
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
1/30.
WB
1/1000. Detects a band of approximately 38, 50 kDa (predicted molecular weight: 47 kDa).
Flow Cyt (Intra)
1/500.

Not suitable with Rat.
追加情報
Is unsuitable for ICC/IF.

ターゲット情報

画像

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NP-I antibody [EPR25683-64] (ab289966)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NP-I antibody [EPR25683-64] (ab289966)

    Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labelling NP-I with ab289966 at 1/2000 followed by LeicaDS9800 (Bond™, Polymer Refine Detection). Positive staining on rat cerebrum. The section was incubated with ab289966 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: PBS instead of primary antibody, followed by LeicaDS9800 (Bond™, Polymer Refine Detection).

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

  • Immunoprecipitation - Anti-NP-I antibody [EPR25683-64] (ab289966)
    Immunoprecipitation - Anti-NP-I antibody [EPR25683-64] (ab289966)

    NP-I was immunoprecipitated from Mouse hypothalamus tissue lysate with ab289966 at 1/30 dilution (2ug in 0.175mg lysates). Western blot was performed on the immunoprecipitate using ab289966 at 1/5000 dilution. Peroxidase IgG Fraction Monoclonal Mouse Anti-Rabbit IgG, light chain specific was used at 1/100000 dilution.

    Lane 1: Mouse hypothalamus tissue lysate 5 ug

    Lane 2: abab289966 IP in Mouse hypothalamus tissue lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab289966 in Mouse hypothalamus tissue lysate

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

    Exposure time: 3.25 seconds

    The 38KDa band might be NPTX1 fragmemts (PMID: 29501530).

  • Flow Cytometry (Intracellular) - Anti-NP-I antibody [EPR25683-64] (ab289966)
    Flow Cytometry (Intracellular) - Anti-NP-I antibody [EPR25683-64] (ab289966)

    Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling NP-I with ab289966 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

  • Western blot - Anti-NP-I antibody [EPR25683-64] (ab289966)
    Western blot - Anti-NP-I antibody [EPR25683-64] (ab289966)
    All lanes : Anti-NP-I antibody [EPR25683-64] (ab289966) at 1/1000 dilution

    Lane 1 : Mouse hypothalamus tissue lysate
    Lane 2 : Mouse brain tissue lysate
    Lane 3 : Rat brain tissue lysate
    Lane 4 : Rat striatum tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 47 kDa
    Observed band size: 38, 50 kDa why is the actual band size different from the predicted?



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

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 29501530)

    Exposure time: 15 seconds.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NP-I antibody [EPR25683-64] (ab289966)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NP-I antibody [EPR25683-64] (ab289966)

    Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labelling NP-I with ab289966 at 1/2000 followed by LeicaDS9800 (Bond™, Polymer Refine Detection) was used. Positive staining on mouse cerebrum. The section was incubated with ab289966 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: PBS instead of primary antibody, followed by LeicaDS9800 (Bond™, Polymer Refine Detection).

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

  • Immunoprecipitation - Anti-NP-I antibody [EPR25683-64] (ab289966)
    Immunoprecipitation - Anti-NP-I antibody [EPR25683-64] (ab289966)

    NP-I was immunoprecipitated from HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab2568364 at 1/30 dilution (2ug in 0.07mg lysates). Western blot was performed on the immunoprecipitate using anti npi antibody epr2568364 immunoprecipitation hela.jpg at 1/5000 dilution. Peroxidase IgG Fraction Monoclonal Mouse Anti-Rabbit IgG, light chain specific was used at 1/100000 dilution.

    Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 2ug

    Lane 2: ab2568364 IP in HeLa whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab289966 in HeLa whole cell lysate

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

    Exposure time: 3.25 seconds

    The 38KDa band might be NPTX1 fragmemts (PMID: 29501530).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NP-I antibody [EPR25683-64] (ab289966)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NP-I antibody [EPR25683-64] (ab289966)

    Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labelling NP-I with ab289966 at 1/2000 followed by LeicaDS9800 (Bond™, Polymer Refine Detection). Positive staining on human cerebrum. The section was incubated with ab289966 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: PBS instead of primary antibody, followed by LeicaDS9800 (Bond™, Polymer Refine Detection).

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

  • Western blot - Anti-NP-I antibody [EPR25683-64] (ab289966)
    Western blot - Anti-NP-I antibody [EPR25683-64] (ab289966)
    All lanes : Anti-NP-I antibody [EPR25683-64] (ab289966) at 1/1000 dilution

    Lane 1 : His-tagged human NPTX1 recombinant protein, 10 ng
    Lane 2 : His-tagged human NPTX2 recombinant protein, 10 ng

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

    Predicted band size: 47 kDa
    Observed band size: 50 kDa why is the actual band size different from the predicted?



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

    Exposure times: Lane 1: 1 seconds; Lane 2: 180 seconds

     

  • Western blot - Anti-NP-I antibody [EPR25683-64] (ab289966)
    Western blot - Anti-NP-I antibody [EPR25683-64] (ab289966)
    All lanes : Anti-NP-I antibody [EPR25683-64] (ab289966) at 1/1000 dilution

    Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate
    Lane 2 : HeLa transfected with siRNA specifically targeti NPTX1 whole cell lysate

    Lysates/proteins at 16 µg per lane.

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

    Predicted band size: 47 kDa
    Observed band size: 50 kDa why is the actual band size different from the predicted?



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

    Exposure time: 10 seconds

     

  • Western blot - Anti-NP-I antibody [EPR25683-64] (ab289966)
    Western blot - Anti-NP-I antibody [EPR25683-64] (ab289966)
    All lanes : Anti-NP-I antibody [EPR25683-64] (ab289966) at 1/1000 dilution

    Lane 1 : Human hypothalamus tissue lysate
    Lane 2 : Human heart tissue lysate
    Lane 3 : Untreated Human hypothalamus tissue lysate
    Lane 4 : Human hypothalamus tissue lysate treated with Protein Deglycosylation MIX II

    Lysates/proteins at 20 µg per lane.

    Secondary
    Lanes 1-2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
    Lanes 3-4 : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

    Predicted band size: 47 kDa
    Observed band size: 47, 50 kDa why is the actual band size different from the predicted?



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

    NPTX1 is a glycosylated protein and can be deglycosylated by Protein Deglycosylation MIX II.

    Negative control: human heart (PMID:7695898)

    Exposure time: 7.75 seconds

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NP-I antibody [EPR25683-64] (ab289966)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NP-I antibody [EPR25683-64] (ab289966)

    Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labelling NP-I with ab289966 at 1/2000 followed by LeicaDS9800 (Bond™, Polymer Refine Detection). Negative control: no staining on human heart. The section was incubated with ab289966 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

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

  • Anti-NP-I antibody [EPR25683-64] (ab289966)
    Anti-NP-I antibody [EPR25683-64] (ab289966)

プロトコール

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

参考文献 (0)

ab289966 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab289966 は論文での使用が確認できていません。

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