Overview

  • Product name

    Anti-Brk/PTK6 antibody [EPR21051-96]
    See all Brk/PTK6 primary antibodies
  • Description

    Rabbit monoclonal [EPR21051-96] to Brk/PTK6
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human Brk/PTK6 aa 1-200. The exact sequence is proprietary.
    Database link: Q13882

  • Positive control

    • WB: T-47D, MCF7, HT-29 and PC-3 whole cell lysates; Human breast cancer and skin lysates. IHC-P: Human breast cancer and colon tissues. Flow Cytometry: T-47D cells. IP: T-47D whole cell lysate.
  • General notes

     This product was previously labelled as Brk

     

    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 ab233392 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 1/500. 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 48 kDa (predicted molecular weight: 52 kDa).
IP 1/30.
Flow Cyt 1/600.

Target

  • Function

    Phosphorylates KHDRBS1, KHDRBS2, KHDRBS3 and STAP2/BKS. May function as an intracellular signal transducer in epithelial tissues. Overexpression in mammary cells leads to mitogenically sensitization to EGF, and results in a partially transformed phenotype. Its presence in the nucleus appears to be linked to suppression of tumor progression.
  • Tissue specificity

    Epithelia-specific. Very high level in colon and high levels in small intestine and prostate, and low levels in some fetal tissues. Expressed at low level in some breast tumors, but not in normal breast. Also found in melanocytes. Not expressed in heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas.
  • Sequence similarities

    Belongs to the protein kinase superfamily. Tyr protein kinase family. BRK/PTK6/SIK subfamily.
    Contains 1 protein kinase domain.
    Contains 1 SH2 domain.
    Contains 1 SH3 domain.
  • Post-translational
    modifications

    Autophosphorylated. The phosphorylation of Tyr-447 may lead to the autoinhibition of the enzyme.
  • Cellular localization

    Cytoplasm. Nucleus. Colocalizes with KHDRBS1, KHDRBS2 or KHDRBS3, within the nucleus. In secretory epithelial cells from prostate adenocarcinoma, nuclear localization is higher in low-grade and lower in high-grade regions of the tumors.
  • Information by UniProt
  • Database links

  • Alternative names

    • Breast tumor kinase antibody
    • Breast tumour kinase antibody
    • Protein tyrosine kinase 6 antibody
    • Protein-tyrosine kinase 6 antibody
    • Ptk6 antibody
    • PTK6_HUMAN antibody
    • Tyrosine protein kinase BRK antibody
    • Tyrosine-protein kinase BRK antibody
    see all

Images

  • All lanes :

    Lane 1 : T-47D (human ductal breast epithelial tumor cell line) whole cell lysate
    Lane 2 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate
    Lane 3 : HT-29 (human colorectal adenocarcinoma cell line) whole cell lysate
    Lane 4 : PC-3 (human prostate adenocarcinoma cell line) whole cell lysate
    Lane 5 : Human skin lysate
    Lane 6 : Human breast cancer 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: 52 kDa
    Observed band size: 48 kDa
    why is the actual band size different from the predicted?



    Exposure time : Lane 1: 15 seconds; Lane 2-6: 3 minutes.

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Brk/PTK6 with ab233392 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic and nuclear staining in surface epithelium of human colon (PMID: 10430081) is observed. Counter stained with hematoxylin.

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

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized T-47D (human ductal breast epithelial tumor cell line) cell line labeling Brk/PTK6 with ab233392 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.

  • Brk/PTK6 was immunoprecipitated from 0.35 mg of T-47D (human ductal breast epithelial tumor cell line) whole cell lysate with ab233392 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab233392 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

    Lane 1: T-47D whole cell lysate 10 µg (Input). 

    Lane 2: ab233392 IP in T-47D whole cell lysate. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab233392 in T-47D whole cell lysate.

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

    Exposure time: 3 minutes.

    IgG heavy chain: PMID: 18781181

  • Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling Brk/PTK6 with ab233392 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic and nuclear staining in and membranous staining on human breast cancer cells is observed. Counter stained with hematoxylin.

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

    Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

References

ab233392 has not yet been referenced specifically in any publications.

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