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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)

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Western blot - Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)
  • Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23222-24] to Eph receptor B4/HTK - BSA and Azide free
  • Suitable for: IHC-P, WB
  • Knockout validated
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Unconjugated

Overview

  • Product name

    Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free
    See all Eph receptor B4/HTK primary antibodies

  • Description

    Rabbit monoclonal [EPR23222-24] to Eph receptor B4/HTK - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, WBmore details
    Unsuitable for: Flow Cyt,ICC/IF or IP

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

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

  • Positive control

    • WB: HEK293T, HT-29, HUVEC, HCT116, MCF7, T47D, PC-3, NIH:OVCAR-3, SK-BR-3, GR-M, 4T1 and NIH/3T3 lysates. IHC-P: Human colon, Human colon cancer and Human breast cancer tissues.

  • General notes

    ab269369 is the carrier-free version of ab254301.

    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.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab269369 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
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
Use at an assay dependent concentration. Detects a band of approximately 120 kDa (predicted molecular weight: 108 kDa).
Notes
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.
WB
Use at an assay dependent concentration. Detects a band of approximately 120 kDa (predicted molecular weight: 108 kDa).
Application notes
Is unsuitable for Flow Cyt,ICC/IF or IP.

Target

  • Function

    Receptor for members of the ephrin-B family. Binds to ephrin-B2. May have a role in events mediating differentiation and development.

  • Tissue specificity

    Abundantly expressed in placenta and in a range of primary tissues and malignant cell lines. Expressed in fetal, but not adult, brain, and in primitive and myeloid, but not lymphoid, hematopoietic cells.

  • Sequence similarities

    Belongs to the protein kinase superfamily. Tyr protein kinase family. Ephrin receptor subfamily.
    Contains 2 fibronectin type-III domains.
    Contains 1 protein kinase domain.
    Contains 1 SAM (sterile alpha motif) domain.

  • Post-translational
    modifications

    Autophosphorylated.

  • Cellular localization

    Membrane.
  • Information by UniProt

  • Database links

    • Alternative names

      • Ephb4 antibody
      • EPHB4_HUMAN antibody
      • Ephrin receptor EphB4 antibody
      • Ephrin type-B receptor 4 antibody
      • Hepatoma Transmembrane Kinase antibody
      • HTK antibody
      • MYK 1 antibody
      • MYK1 antibody
      • TYRO 11 antibody
      • TYRO11 antibody
      • Tyrosine-protein kinase receptor HTK antibody
      • Tyrosine-protein kinase TYRO11 antibody
      see all

    Images

    • Western blot - Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)
      Western blot - Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)
      All lanes : Anti-Eph receptor B4/HTK antibody [EPR23222-24] (ab254301) at 1/1000 dilution

      Lane 1 : Wild-type HEK293T cell lysate
      Lane 2 : EPHB4 knockout HEK293T cell lysate
      Lane 3 : MCF7 cell lysate
      Lane 4 : HT-29 cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

      Predicted band size: 108 kDa
      Observed band size: 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 (ab254301).

      Lanes 1-4: Merged signal (red and green). Green - ab254301 observed at 125 kDa. Red - loading control ab8245 observed at 36 kDa.

       ab254301 Anti-Eph receptor B4/HTK antibody [EPR23222-24] was shown to specifically react with Eph receptor B4/HTK in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266733 (knockout cell lysate ab257429) was used. Wild-type and Eph receptor B4/HTK knockout samples were subjected to SDS-PAGE. ab254301 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)

      Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling Eph receptor B4 with ab254301 at 1/1000 dilution (0.545ug/ml) followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at a ready to use dilution. Mainly membranous staining in cancer cells of human breast cancer (PMID: 16816380) is observed. The section was incubated with ab255611 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at Ready to use dilution.

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254301).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)

      Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labeling Eph receptor B4 with ab254301 at 1/1000 dilution (0.545ug/ml) followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at a ready to use dilution. Positive staining in cancer cells of human colon cancer (PMID: 19366806) is observed. The section was incubated with ab255611 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at Ready to use dilution.

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254301).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)

      Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Eph receptor B4 with ab254301 at 1/1000 dilution (0.545ug/ml) followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at a ready to use dilution. Weak positive staining in base of crypts of human colon (PMID:19366806) is observed. The section was incubated with ab255611 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at Ready to use dilution.

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254301).

    • Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)
      Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)

    Protocols

    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

    References (0)

    Publishing research using ab269369? Please let us know so that we can cite the reference in this datasheet.

    ab269369 has not yet been referenced specifically in any publications.

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