Overview

  • Product name

    Anti-HOXB4 antibody [EP1919Y]
    See all HOXB4 primary antibodies
  • Description

    Rabbit monoclonal [EP1919Y] to HOXB4
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, ICC, Flow Cyt, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human HOXB4 aa 100-200 (internal sequence). The exact sequence is proprietary.

  • Positive control

    • WB: K-562 lysates. IHC-P: human lung carcinoma tissue. ICC/IF: K-562 cells. FC: K-562 cells. IP: K-562 cells.
  • General notes

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

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab76093 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
WB 1/1000 - 1/10000. Predicted molecular weight: 28 kDa.
IP 1/60.

For unpurified use please use at 1/40

ICC 1/100 - 1/250.
Flow Cyt 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols

Target

  • Function

    Sequence-specific transcription factor which is part of a developmental regulatory system that provides cells with specific positional identities on the anterior-posterior axis.
  • Sequence similarities

    Belongs to the Antp homeobox family. Deformed subfamily.
    Contains 1 homeobox DNA-binding domain.
  • Developmental stage

    Expressed in whole embryos and fetuses at 5-9 weeks from conception.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • Homeo box 2F antibody
    • Homeo box B4 antibody
    • Homeobox 2F antibody
    • Homeobox B4 antibody
    • Homeobox protein Hox B4 antibody
    • Homeobox protein Hox-2.6 antibody
    • Homeobox protein Hox-2F antibody
    • Homeobox protein Hox-B4 antibody
    • Homeobox protein HoxB4 antibody
    • HOX 2 antibody
    • Hox 2.6 antibody
    • Hox 2F antibody
    • HOX B4 antibody
    • HOX2 antibody
    • Hox2.6 antibody
    • Hox2F antibody
    • HOXB 4 antibody
    • hoxb4 antibody
    • HXB4_HUMAN antibody
    see all

Images

  • Immunocytochemistry/ Immunofluorescence analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labeling HOXB4 with purified ab76093 at 1:100 dilution (10 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
  • ab76093 (Purified) at 1:60 dilution (2 µg) immunoprecipitating HOXB4 in K-562 whole cell lysate.
    Lane 1 (input): K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate 10 µg
    Lane 2 (+): ab76093 & K-562 whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76093 in K-562 whole cell lysate
    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.
  • Flow Cytometry analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labeling HOXB4 with purified ab76093 at 1:100 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue sections labeling HOXB4 with purified ab76093 at 1:100 dilution (11.86 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
  • Anti-HOXB4 antibody [EP1919Y] (ab76093) at 1/1000 dilution (Purified) + K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates at 15 µg

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

    Predicted band size: 28 kDa
    Observed band size: 34 kDa
    why is the actual band size different from the predicted?

  • Immunohistochemical analysis of paraffin-embedded human placental tissue, using unpurified ab76093, at 1/100 dilution.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Overlay histogram showing K562 cells stained with unpurified ab76093 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76093, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References

This product has been referenced in:

  • Zhou G  et al. Homeobox B4 inhibits breast cancer cell migration by directly binding to StAR-related lipid transfer domain protein 13. Oncol Lett 14:4625-4632 (2017). Read more (PubMed: 29085460) »
See 1 Publication for this product

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