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リコンビナントRabMAb

Anti-BTK (phospho Y551) 抗体 [EP267Y] (ab40770)

Submit a review Submit a question References (8)
Indirect ELISA - Anti-BTK (phospho Y551) antibody [EP267Y] (ab40770)
  • Dot Blot - Anti-BTK (phospho Y551) antibody [EP267Y] (ab40770)
  • Western blot - Anti-BTK (phospho Y551) antibody [EP267Y] (ab40770)
  • Western blot - Anti-BTK (phospho Y551) antibody [EP267Y] (ab40770)
  • Immunocytochemistry/ Immunofluorescence - Anti-BTK (phospho Y551) antibody [EP267Y] (ab40770)
  • Anti-BTK (phospho Y551) antibody [EP267Y] (ab40770)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP267Y] to BTK (phospho Y551)
  • Suitable for: Flow Cyt (Intra), ICC/IF, Indirect ELISA, WB, Dot blot
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

製品の概要

  • 製品名

    Anti-BTK (phospho Y551) antibody [EP267Y]
    BTK 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [EP267Y] to BTK (phospho Y551)

  • 由来種

    Rabbit

  • 特異性

    Stimulation may be required to allow detection of the phosphorylated protein.  Please see images below for recommended treatment conditions and positive controls.

    ab40770 also reacts with family proteins including BMX (phospho Y566), ITK (phospho Y512), TEC (phospho Y519) and TXK (phospho Y420).

  • アプリケーション

    適用あり: Flow Cyt (Intra), ICC/IF, Indirect ELISA, WB, Dot blotmore details
    適用なし: Flow Cyt or IP

  • 種交差性

    交差種: Human

  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • ポジティブ･コントロール

    • WB: Ramos treated with 1mM pervanadate whole cell lysates, K-562 treated with 1mM pervanadate whole cell lysate ICC/IF: Ramos cells

  • 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.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

製品の特性

  • 製品の状態

    Liquid

  • 保存方法

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.

  • バッファー

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 50% Glycerol (glycerin, glycerine), 0.05% BSA, 49% PBS
  • 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

    EP267Y

  • 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ab40770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
Flow Cyt (Intra)
Use at an assay dependent concentration.
ICC/IF
1/500.
Indirect ELISA
Use at an assay dependent concentration.
WB
1/2000. Detects a band of approximately 74 kDa (predicted molecular weight: 74 kDa).
Dot blot
1/1000.
特記事項
Flow Cyt (Intra)
Use at an assay dependent concentration.
ICC/IF
1/500.
Indirect ELISA
Use at an assay dependent concentration.
WB
1/2000. Detects a band of approximately 74 kDa (predicted molecular weight: 74 kDa).
Dot blot
1/1000.
追加情報
Is unsuitable for Flow Cyt or IP.

ターゲット情報

  • 機能

    Plays a crucial role in B-cell ontogeny. Transiently phosphorylates GTF2I on tyrosine residues in response to B-cell receptor cross-linking. Required for the formation of functional ARID3A DNA-binding complexes.

  • 関連疾患

    Defects in BTK are the cause of X-linked agammaglobulinemia (XLA) [MIM:300755]; also known as X-linked agammaglobulinemia type 1 (AGMX1) or immunodeficiency type 1 (IMD1). XLA is a humoral immunodeficiency disease which results in developmental defects in the maturation pathway of B-cells. Affected boys have normal levels of pre-B-cells in their bone marrow but virtually no circulating mature B-lymphocytes. This results in a lack of immunoglobulins of all classes and leads to recurrent bacterial infections like otitis, conjunctivitis, dermatitis, sinusitis in the first few years of life, or even some patients present overwhelming sepsis or meningitis, resulting in death in a few hours. Treatment in most cases is by infusion of intravenous immunoglobulin.
    Defects in BTK may be the cause of X-linked hypogammaglobulinemia and isolated growth hormone deficiency (XLA-IGHD) [MIM:307200]; also known as agammaglobulinemia and isolated growth hormone deficiency or Fleisher syndrome or isolated growth hormone deficiency type 3 (IGHD3). In rare cases XLA is inherited together with isolated growth hormone deficiency (IGHD).

  • 配列類似性

    Belongs to the protein kinase superfamily. Tyr protein kinase family. TEC subfamily.
    Contains 1 Btk-type zinc finger.
    Contains 1 PH domain.
    Contains 1 protein kinase domain.
    Contains 1 SH2 domain.
    Contains 1 SH3 domain.

  • 翻訳後修飾

    Autophosphorylated on Tyr-223 and Tyr-551. Phosphorylation of Tyr-223 may create a docking site for a SH2 containing protein.

  • 細胞内局在

    Cytoplasm. Membrane. Nucleus.
  • Information by UniProt

  • 参照データベース

    • 別名

      • Agammaglobulinaemia tyrosine kinase antibody
      • AGMX 1 antibody
      • AGMX1 antibody
      • AT antibody
      • ATK antibody
      • B cell progenitor kinase antibody
      • B-cell progenitor kinase antibody
      • BPK antibody
      • Bruton agammaglobulinemia tyrosine kinase antibody
      • Bruton tyrosine kinase antibody
      • Bruton’s Tyrosine Kinase antibody
      • Btk antibody
      • BTK_HUMAN antibody
      • dominant-negative kinase-deficient Brutons tyrosine kinase antibody
      • IMD 1 antibody
      • IMD1 antibody
      • MGC126261 antibody
      • MGC126262 antibody
      • OTTHUMP00000063593 antibody
      • PSCTK 1 antibody
      • PSCTK1 antibody
      • truncated Bruton agammaglobulinemia tyrosine kinase antibody
      • Tyrosine protein kinase BTK antibody
      • Tyrosine-protein kinase BTK antibody
      • tyrosine-protein kinase BTK isoform (lacking exon 14 antibody
      • XLA antibody
      see all

    画像

    • Indirect ELISA - Anti-BTK (phospho Y551) antibody [EP267Y] (ab40770)
      Indirect ELISA - Anti-BTK (phospho Y551) antibody [EP267Y] (ab40770)

      ELISA using ab40770 at varying antibody concentrations and antigen concentration of 1000 ng/ml. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) (1/2500) was used as the secondary antibody. Substrate solution was p-nitrophenyl phosphate (PNPP).

    • Dot Blot - Anti-BTK (phospho Y551) antibody [EP267Y] (ab40770)
      Dot Blot - Anti-BTK (phospho Y551) antibody [EP267Y] (ab40770)

      Dot blot analysis of Human BTK non-phospho peptide (lane 1), Human BTK Y551 phospho peptide (lane 2), Human BMX non-phospho peptide (lane 3), Human BMX Y566 phospho peptide (lane 4), Human ITK non-phospho peptide (lane 5), Human ITK Y512 phospho peptide (lane 6), Human TEC non-phospho peptide (lane 7), Human TEC Y519 phospho peptide (lane 8), Human TXK non-phospho peptide (lane 9) and Human TXK Y420 phospho peptide (lane 10) with ab40770 at a dilution of 1/1000. ab97051 Peroxidase conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody at a dilution of 1/20,000.

      Blocking buffer and concentration : 5% NFDM/TBST

      Diluting buffer and concentration : 5% NFDM/TBST

      Exposure time: 180 seconds

       

       

    • Western blot - Anti-BTK (phospho Y551) antibody [EP267Y] (ab40770)
      Western blot - Anti-BTK (phospho Y551) antibody [EP267Y] (ab40770)
      All lanes : Anti-BTK (phospho Y551) antibody [EP267Y] (ab40770) at 1/1000 dilution

      Lane 1 : Untreated K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate
      Lane 2 : K-562 treated with 1mM pervanadate for 30 min whole cell lysate
      Lane 3 : K-562 treated with 1mM pervanadate for 30 min whole cell lysate, then the membrane treated with Alkaline Phosphatase for 1 hour

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 74 kDa
      Observed band size: 75 kDa why is the actual band size different from the predicted?


      Exposure time: 180 seconds


      Blocking buffer and concentration : 5% NFDM/TBST

      Diluting buffer and concentration : 5% NFDM/TBST

       

    • Western blot - Anti-BTK (phospho Y551) antibody [EP267Y] (ab40770)
      Western blot - Anti-BTK (phospho Y551) antibody [EP267Y] (ab40770)
      All lanes : Anti-BTK (phospho Y551) antibody [EP267Y] (ab40770) at 1/1000 dilution

      Lane 1 : Untreated Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysate
      Lane 2 : Ramos treated with 1mM pervanadate for 30 min whole cell lysate
      Lane 3 : Ramos treated with 1mM pervanadate for 30 min whole cell lysate, then the membrane treated with Alkaline Phosphatase for 1 hour

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 74 kDa
      Observed band size: 75 kDa why is the actual band size different from the predicted?


      Exposure time: 7 seconds


      Blocking buffer and concentration : 5% NFDM/TBST

      Diluting buffer and concentration : 5% NFDM/TBST

       

    • Immunocytochemistry/ Immunofluorescence - Anti-BTK (phospho Y551) antibody [EP267Y] (ab40770)
      Immunocytochemistry/ Immunofluorescence - Anti-BTK (phospho Y551) antibody [EP267Y] (ab40770)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Ramos (human Burkitt's lymphoma B lymphocyte) cells labelling BTK with primary antibody anti- BTK (phospho Y551) (ab40770) at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150081) secondary antibody at 1/1000 dilution (2.0 µg/mL). Confocal image showing increased cytoplasmic staining in Ramos cells treated with pervanadate (1 mM) for 30 min and then the signal decreased after Alkaline Phosphatase treatment 37℃ for 1 hour. Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (ab195889) was used to counterstain tubulin at 1/200 dilution (2.5 µg/mL). The nuclear counter stain is DAPI (blue).

      The secondary antibody only control is : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (2.0 µg/mL).

    • Anti-BTK (phospho Y551) antibody [EP267Y] (ab40770)
      Anti-BTK (phospho Y551) antibody [EP267Y] (ab40770)

    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 (8)

    ab40770 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。

    ab40770 は 8 報の論文で使用されています。

    • Sonowal H  et al. Luxeptinib interferes with LYN-mediated activation of SYK and modulates BCR signaling in lymphoma. PLoS One 18:e0277003 (2023). PubMed: 36888611
    • Aryal M  et al. The HIV-1 protein Nef activates the Tec family kinase Btk by stabilizing an intermolecular SH3-SH2 domain interaction. Sci Signal 15:eabn8359 (2022). PubMed: 36126115
    • Shu ST  et al. Visualization of Host Cell Kinase Activation by Viral Proteins Using GFP Fluorescence Complementation and Immunofluorescence Microscopy. Bio Protoc 11:e4068 (2021). PubMed: 34327265
    • Li WF  et al. HIV-1 Nef dimers short-circuit immune receptor signaling by activating Tec-family kinases at the host cell membrane. J Biol Chem 295:5163-5174 (2020). PubMed: 32144207
    • Mo ZQ  et al. Molecular characterization and function analysis of grouper (Epinephelus coioides) Bruton's tyrosine kinase BTK. Fish Shellfish Immunol 77:91-99 (2018). WB . PubMed: 29567142
    • Rushworth SA  et al. Identification of Bruton's tyrosine kinase as a therapeutic target in acute myeloid leukemia. Blood 123:1229-38 (2014). Human . PubMed: 24307721
    • Daniel JL  et al. Cbl-b is a novel physiologic regulator of glycoprotein VI-dependent platelet activation. J Biol Chem 285:17282-91 (2010). PubMed: 20400514
    • Grasis JA  et al. In vivo significance of ITK-SLP-76 interaction in cytokine production. Mol Cell Biol 30:3596-609 (2010). PubMed: 20457812

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