Overview

  • Product name

    Anti-BTK antibody [EPR20445]
    See all BTK primary antibodies
  • Description

    Rabbit monoclonal [EPR20445] to BTK
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WB, ICC/IF, IP, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human BTK aa 350 to the C-terminus. The exact sequence is proprietary.
    Database link: Q06187

  • Positive control

    • WB: Ramos and NAMALWA whole cell lysates; Human lymph node lysate. IHC-P: Human tonsil and colon tissues. ICC/IF: Ramos cells. Flow Cyt: Ramos cells. IP: Ramos whole cell lysate.
  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab208937 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 76 kDa (predicted molecular weight: 76 kDa).
ICC/IF 1/100.
IP 1/30.
Flow Cyt 1/600.

Target

  • Function

    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.
  • Involvement in disease

    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).
  • Sequence similarities

    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.
  • Post-translational
    modifications

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

    Cytoplasm. Membrane. Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • 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

Images

  • All lanes : Anti-BTK antibody [EPR20445] (ab208937) at 1/1000 dilution

    Lane 1 : Ramos (human Burkitt's lymphoma cell line) whole cell lysate
    Lane 2 : NAMALWA (human Burkitt's lymphoma B lymphocyte) whole cell lysate
    Lane 3 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

    Developed using the ECL technique.

    Predicted band size: 76 kDa
    Observed band size: 76 kDa


    Exposure time: 32 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The Jurkat T cell line is negative for BTK (PMID 24759210).

    This image is performed on a BIO-RAD® ChemiDoc™ MP instrument.

     

  • Anti-BTK antibody [EPR20445] (ab208937) at 1/1000 dilution + Human lymph node lysate at 10 µg

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

    Developed using the ECL technique.

    Predicted band size: 76 kDa
    Observed band size: 76 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    This image is performed on a BIO-RAD® ChemiDoc™ MP instrument.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling BTK with ab208937 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Strong cytoplasmic staining in mantle zone and weaker cytoplasmic staining in the germinal center of human tonsil (PMID: 25433814). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded human colon tissue labeling BTK with ab208937 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Strong cytoplasmic staining on lymphoid nodule in human colon (PMID: 25433814). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunofluorescent analysis of 100% methanol-fixed Ramos (human Burkitt's lymphoma cell line) and Jurkat (human T cell leukemia cell line from peripheral blood) cells labeling BTK with ab208937 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing cytoplasmic staining on Ramos cell line.

    Negative control: Jurkat cell line (PMID: 24759210).

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized Jurkat (human T cell leukemia cell line from peripheral blood) (left panel) and Ramos (human Burkitt's lymphoma cell line) (right panel) cell lines labeling BTK with ab208937 at 1/600 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

    Negative control: Jurkat cell line (PMID: 24759210).

  • BTK was immunoprecipitated from 0.35 mg of Ramos (human Burkitt's lymphoma cell line) whole cell lysate with ab208937 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab208937 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: Ramos whole cell lysate 10 µg (Input). 

    Lane 2: ab208937 IP in Ramos whole cell lysate. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab208937 in Ramos whole cell lysate.

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

    Exposure time: 3 seconds.

References

This product has been referenced in:

  • Wang D  et al. Bone Marrow Molecular Markers Associated with Relapsed/Refractory Activated B-Cell-Like Diffuse Large B-Cell Lymphoma. Biomed Res Int 2018:1042597 (2018). Read more (PubMed: 30534553) »
See 1 Publication for this product

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